LOCUS       pCS2+_ALFAgrad          5589 bp    DNA     circular UNA 11-MAY-2026
DEFINITION  19.
ACCESSION   urn.local...rm-klr9txw
VERSION     urn.local...rm-klr9txw
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     regulatory      34..51
                     /note="Geneious type: Promoter"
                     /standard_name="Sp6 prom"
     5'UTR           109..151
                     /note="Xenopus globin 5′ UTR sequence"
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="Xenopus globin 5′ UTR"
     gene            167..817
                     /standard_name="zebrafish fbxw11b"
     gene            818..1192
                     /standard_name="NbALFA"
     3'UTR           1202..1524
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="prrg2 3'UTR"
     regulatory      complement(1615..1633)
                     /note="Geneious type: Promoter"
                     /standard_name="T7 prom"
     regulatory      1651..1772
                     /note="Geneious type: terminator"
                     /note="simian vacuolating virus 40 (SV40) polyA signal"
                     /regulatory_class="terminator"
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="SV40 polyA"
     misc_feature    complement(1798..1830)
                     /note="KT3 epitope tag used for protein detection and
                     purification"
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on protein similarity"
                     /standard_name="KT3"
     regulatory      complement(1884..1903)
                     /note="Geneious type: Promoter"
                     /standard_name="T3 prom"
     misc_feature    complement(1942..1969)
                     /note="Geneious type: Regulatory Sequence"
                     /standard_name="lacO reg"
     misc_feature    1948..1964
                     /note="Lac operon operator"
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="lacO"
     regulatory      complement(1973..2002)
                     /note="Geneious type: Promoter"
                     /standard_name="lac prom"
     misc_feature    1980..1991
                     /note="C-tag used for protein purification"
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on protein similarity"
                     /standard_name="C"
     regulatory      2017..2038
                     /note="catabolite activator protein binding site"
                     /regulatory_class="other"
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="CAP BS"
     rep_origin      complement(2326..2914)
     CDS             complement(3085..3945)
                     /note="confers ampicillin resistance"
                     /product="beta-lactamase"
                     /gene="bla"
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="ampR"
     regulatory      complement(3946..4050)
                     /note="Geneious type: promoter"
                     /note="Promoter for ampicillin resistance gene
                     (b-lactamase)"
                     /regulatory_class="promoter"
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="amp"
     rep_origin      complement(4076..4531)
     regulatory      4603..5589
                     /note="Geneious type: promoter"
                     /note="Human cytomegalovirus (CMV) immediate early
                     promoter"
                     /regulatory_class="promoter"
                     /note="Derived using Geneious Prime 2026.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="CMV IE94"
ORIGIN      
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcaga atcccatcga ttcgaattca agagcttgct tgttcttttt
      121 gcagaagctc agaataaacg ctcaactttg gcagatcgcc gccaccatgg agacggagat
      181 ggaggacaaa accttagagc agatgaacac ctctgtcatg gacccgcaga ctgcagaccg
      241 ctctccaaag atcacattga ttaagtctac ctttatctgt cctcaggtga gtaatggacc
      301 cttgacgggc tctaggaagc ggccgtcaga gggaaattac gagaaggaaa aggacgtgtg
      361 catccagctc tttgatcaat ggtctgaggc cgaccaggtg gagtttgtgg aacacctgat
      421 ctcacgcatg tgccattacc aacacggcca cataaactcc tacctcaaac ccatgctcca
      481 aagggatttc attacggcac tgccagctca gggtctggat catatagcgg agaatatctt
      541 gtcgttcctg gatgctcgct cgttgtgttc ggctgagctg gtgtgtaagg aatggcagcg
      601 tgtcatatct gaaggcatgt tgtggaagaa gctgattgag aggatggttc ggacagaccc
      661 gctgtggaaa ggcctatcag agagacacca gtgggaaaaa tacctgttca agaaccgcac
      721 aacagaagtt cctccaaact cgtactaccg ctctctttat ccaaaaatca ttcaggacat
      781 agagaccatt gaagcaaact ggaggtgtgg tcgacatatg gatgaagtcc agctgcagga
      841 gtccggtggt ggcctggttc aaccaggtgg ctccctacga ctgtcctgta ccgcatccgg
      901 agttacgatt agcgctctga atgctatggc aatgggatgg tatcgtcagg caccgggaga
      961 gcgtcgtgtc atggtagctg ctgtctctga acgtggaaat gctatgtatc gtgaatcagt
     1021 tcagggccgt tttaccgtta cccgcgattt caccaacaaa atggtcagtc tgcagatgga
     1081 caacctgaaa ccggaggata cagcggtgta ctactgccat gtcctggaag atcgcgttga
     1141 tagcttccac gattattggg gacaggggac ccaggtcact gtgtcctcat aataagagct
     1201 ctctcaaaat gaggaatgag caatgatttt ttaaagaaaa attttacgag ttggatttta
     1261 tttcactggt gaacggagtg cgcggctaca ttgaggcttc gagatggtgg tcatgcgcca
     1321 cagacaaatg tagtttcagt cactgtttgg ctaacttatt ccacttgggc tgtctgttta
     1381 gcggcttgcc ttgtttatta cattattttg ctaccaaagg ttttattgtg aattgccaat
     1441 gttttcatga atcaagtttg cgtcatcttt aaatagttgt ttagacattt taaatgttca
     1501 ttgttaataa aaataactaa acaactagaa tacatacttc taatcattcc atagctaata
     1561 catacttcta atcattccat agctaataca tacttctaat cattccatac tagaactata
     1621 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca
     1681 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1741 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1801 ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt
     1861 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     1921 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1981 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2041 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2101 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2161 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2761 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3061 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3121 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3181 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3241 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3301 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3361 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3421 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3481 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3541 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3601 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3661 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3721 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3781 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3841 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3901 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3961 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4021 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     4081 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4141 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4201 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4261 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4321 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4381 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4441 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4501 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     4561 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat agccaattca
     4621 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
     4681 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca
     4741 atatggcgga cttggcacca tgccaattca atatggcgga cttggcactg tgccaactgg
     4801 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc
     4861 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc
     4921 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata
     4981 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat
     5041 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag
     5101 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg
     5161 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg
     5221 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc
     5281 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc
     5341 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag
     5401 gacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat
     5461 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt
     5521 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt
     5581 ttagggaac
//
