LOCUS       pCS2+_Halo-NbALFA_boomUTR        5859 bp    DNA     circular UNA 11-MAY-2026
DEFINITION  18.
ACCESSION   urn.local...qj-klr9qcb
VERSION     urn.local...qj-klr9qcb
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    1..166
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="pCS2+eGFP-OptZF_NbALFA boomUTR PCR Product
                     - pCS2+eGFP+ prrg2 3’UTR PCR Product (concatenated
                     sequence 1) (concatenated sequence 1)"
     regulatory      34..51
                     /note="Geneious type: Promoter"
                     /standard_name="Sp6 prom"
     primer_bind     complement(147..166)
                     /standard_name="Nb Nterm R"
     misc_feature    152..166
                     /note="Geneious type: ligation"
                     /standard_name="Ligation"
     misc_feature    152..1078
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="Halo (concatenated sequence 2)"
     misc_feature    167..169
                     /note="Geneious type: Editing History Insertion"
                     /standard_name
     misc_feature    <170..>1063
                     /note="HaloTag used for protein expression and
                     purification"
                     /product="haloalkane dehalogenase"
                     /gene="HT2"
                     /note="Derived using Geneious Prime 2020.1.1 'Annotate
                     from Database' based on protein similarity"
                     /standard_name="HaloTag®"
     primer_bind     complement(1045..1063)
                     /standard_name="Halo (897 R) - pCS2+eGFP-OptZF_NbALFA
                     boomUTR PCR Product"
     misc_feature    1063^1064
                     /note="Geneious type: Editing History Deletion"
                     /standard_name="TAA"
     misc_feature    1064..1078
                     /note="Geneious type: ligation"
                     /standard_name="Ligation"
     misc_feature    1064..5859
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="pCS2+eGFP-OptZF_NbALFA boomUTR PCR Product
                     extraction (concatenated sequence 3)"
     misc_feature    <1064..1481
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="pCS2+zGrad OptZF_NbALFA PCR Product
                     (concatenated sequence 2)"
     misc_feature    join(1451..>1462,<1064..1084)
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="pCS2+zGrad PCR Product (concatenated
                     sequence 1)"
     primer_bind     1064..1083
                     /standard_name="pCS2+zGrad OptZF_NbALFA PCR Product (1 F)
                     - pCS2+eGFP+ prrg2 3’UTR PCR Product"
     misc_feature    1068..>1462
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="OptZF_NbALFA (concatenated sequence 2)"
     gene            1079..1450
                     /standard_name="optZF_NbALFA"
     primer_bind     complement(1437..1462)
                     /standard_name="pCS2+zGrad OptZF_NbALFA PCR Product (399
                     R) - pCS2+eGFP+ prrg2 3’UTR PCR Product"
     CDS             <1451..1453
                     /codon_start=1
                     /product="GFP-binding fragment of a single-chain camelid 
                     antibody (Rothbauer et al., 2008)"
                     /translation
                     /standard_name="GFP nanobody"
     misc_feature    1451..>1462
                     /note="Geneious type: ligation"
                     /standard_name="Ligation"
     misc_feature    1460..>1462
                     /note="Geneious type: polyA_signal"
                     /note="SV40 polyadenylation signal"
                     /standard_name="SV40 poly(A) signal"
     misc_feature    1463..1481
                     /note="Geneious type: ligation"
                     /standard_name="Ligation"
     misc_feature    1463..>166
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="pCS2+eGFP+ prrg2 3’UTR PCR Product
                     (concatenated sequence 1)"
     regulatory      complement(1885..1903)
                     /note="Geneious type: Promoter"
                     /standard_name="T7 prom"
     regulatory      complement(2154..2173)
                     /note="Geneious type: Promoter"
                     /standard_name="T3 prom"
     misc_feature    complement(2212..2239)
                     /note="Geneious type: Regulatory Sequence"
                     /standard_name="lacO reg"
     regulatory      complement(2243..2272)
                     /note="Geneious type: Promoter"
                     /standard_name="lac prom"
     misc_feature    complement(2581..3200)
                     /note="Geneious type: Origin of Replication"
                     /standard_name="pBR322 origin"
     misc_feature    complement(3355..4215)
                     /note="Geneious type: Marker"
                     /standard_name="amp marker"
     regulatory      complement(4257..4285)
                     /note="Geneious type: Promoter"
                     /standard_name="amp prom"
     misc_feature    complement(4478..4784)
                     /note="Geneious type: Origin of Replication"
                     /standard_name="f1 origin"
ORIGIN      
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcaga atcccatcga ttcgaattca agagcttgct tgttcttttt
      121 gcagaagctc agaataaacg ctcaactttg gcagatcgcc gccaccatgg cagaaatcgg
      181 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
      241 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
      301 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
      361 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
      421 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
      481 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
      541 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
      601 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
      661 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
      721 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
      781 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
      841 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
      901 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
      961 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1021 gatcgcgcgc tggctgtcga cgctcgagat ttccggcctc gagaggtgtg gtcgacatat
     1081 ggatgaagtc cagctgcagg agtccggtgg tggcctggtt caaccaggtg gctccctacg
     1141 actgtcctgt accgcatccg gagttacgat tagcgctctg aatgctatgg caatgggatg
     1201 gtatcgtcag gcaccgggag agcgtcgtgt catggtagct gctgtctctg aacgtggaaa
     1261 tgctatgtat cgtgaatcag ttcagggccg ttttaccgtt acccgcgatt tcaccaacaa
     1321 aatggtcagt ctgcagatgg acaacctgaa accggaggat acagcggtgt actactgcca
     1381 tgtcctggaa gatcgcgttg atagcttcca cgattattgg ggacagggga cccaggtcac
     1441 tgtgtcctca taatacgtag attaagagct ctctcaaaat gaggaatgag caatgatttt
     1501 ttaaagaaaa attttacgag ttggatttta tttcactggt gaacggagtg cgcggctaca
     1561 ttgaggcttc gagatggtgg tcatgcgcca cagacaaatg tagtttcagt cactgtttgg
     1621 ctaacttatt ccacttgggc tgtctgttta gcggcttgcc ttgtttatta cattattttg
     1681 ctaccaaagg ttttattgtg aattgccaat gttttcatga atcaagtttg cgtcatcttt
     1741 aaatagttgt ttagacattt taaatgttca ttgttaataa aaataactaa acaactagaa
     1801 tacatacttc taatcattcc atagctaata catacttcta atcattccat agctaataca
     1861 tacttctaat cattccatac tagaactata gtgagtcgta ttacgtagat ccagacatga
     1921 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1981 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2041 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2101 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     2161 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2221 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2281 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2341 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2401 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2461 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2521 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2581 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2641 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2701 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2761 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2821 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2881 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2941 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3001 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3061 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3121 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3181 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3241 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3301 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3361 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3421 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3481 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3541 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3601 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3661 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3721 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3781 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3841 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3901 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3961 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4021 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4081 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4141 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4201 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4261 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4321 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     4381 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4441 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4501 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4561 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4621 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4681 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4741 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4801 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4861 ctattacgcc agtcgaccat agccaattca atatggcgta tatggactca tgccaattca
     4921 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     4981 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
     5041 atatggcgga cttggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
     5101 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
     5161 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
     5221 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
     5281 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
     5341 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
     5401 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
     5461 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
     5521 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
     5581 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
     5641 gtaacttggc aagtacatta ctattggaag gacgccaggg tacattggca gtactcccat
     5701 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
     5761 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
     5821 taatgggagg tctatataag caatgctcgt ttagggaac
//
