LOCUS       sequence-311681-        3096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3096)
  AUTHORS   Nam HS, Benezra R
  TITLE     High levels of Id1 expression define B1 type adult neural stem 
            cells.
  JOURNAL   Cell Stem Cell. 2009 Nov 6. 5(5):515-26.
  PUBMED    19896442
REFERENCE   2  (bases 1 to 3096)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Stem 
            Cell. 2009 Nov 6. 5(5):515-26."
COMMENT     Sequence Label: pKO2.2
FEATURES             Location/Qualifiers
     source          1..3096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             262..834
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     intron          911..976
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    1037..1171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1074..1093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1128..1147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1191..1207
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1277..1286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        1330..1434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1435..1503,1504..2295)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1653..1672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2466..3054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2955..2974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggaccgaac aactccgccg cgcgcgctcc gtgtaggccg gtgcgggcgg ccccgtagcg
       61 caagggaggg cgggaaagga aggggcggga cacaagggcg aatctataaa gggcgtcact
      121 cagccagttc tctcctcaga agcgccgaga gcgcgaccgg gacggttgga gaagaaggtg
      181 gctcccggaa gggggagaga caaactgccg taacctctgc cgttcaggat ctgcgacctg
      241 caggtcctcg ccatggaccc tgatgatgtt gttgattctt ctaaatcttt tgtgatggaa
      301 aacttttctt cgtaccacgg gactaaacct ggttatgtag attccattca aaaaggtata
      361 caaaagccaa aatctggtac acaaggaaat tatgacgatg attggaaagg gttttatagt
      421 accgacaata aatacgacgc tgcgggatac tctgtagata atgaaaaccc gctctctgga
      481 aaagctggag gcgtggtcaa agtgacgtat ccaggactga cgaaggttct cgcactaaaa
      541 gtggataatg ccgaaactat taagaaagag ttaggtttaa gtctcactga accgttgatg
      601 gagcaagtcg gaacggaaga gtttatcaaa aggttcggtg atggtgcttc gcgtgtagtg
      661 ctcagccttc ccttcgctga ggggagttct agcgttgaat atattaataa ctgggaacag
      721 gcgaaagcgt taagcgtaga acttgagatt aattttgaaa cccgtggaaa acgtggccaa
      781 gatgcgatgt atgagtatat ggctcaagcc tgtgcaggaa atcgtgtcag gcgatctctt
      841 tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta
      901 aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat
      961 tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc
     1021 agatccacta ggatctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1081 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1141 actcatcaat gtatcttatc atgtctggat cgtagttcta gagcggaccg gtaaaacgac
     1201 ggccagtgtt aactctagga attctctagg gtacctctag gagatcttcg ctaccttagg
     1261 accgttatag ttagcggccg ccatggtcat agctgtttga cgtcaggtgg cacttttcgg
     1321 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1381 ctcacgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1441 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1501 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1561 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1621 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1681 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1741 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1801 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1861 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1921 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1981 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2041 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2101 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2161 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2221 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2281 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2341 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2401 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2461 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2521 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2581 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2641 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2701 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2761 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2821 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2881 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2941 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3001 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3061 agcaacgcgg cctttttacg gttctggcct tttgca
//
