LOCUS       sequence-14381-e        3310 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 3310)
  AUTHORS   Dmitriev PV, Vassetzky YS
  TITLE     A set of vectors for introduction of antibiotic resistance genes by 
            in vitro Cre-mediated recombination.
  JOURNAL   BMC Res Notes. 2008 Dec 23;1:135. doi: 10.1186/1756-0500-1-135.
  PUBMED    19105832
REFERENCE   2  (bases 1 to 3310)
  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: "BMC Res 
            Notes."; date: "2008-12-23"; volume: "1:135. doi"; pages: " 
            10.1186/1756-0500-1-135"
COMMENT     Sequence Label: pINS-Puro
FEATURES             Location/Qualifiers
     source          1..3310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(25..44)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     79..98
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(365..964)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(448..468)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     945..964
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1057..1386)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1071..1206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1371..1391
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(1407..1423)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1408..1424)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(1668..2327)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2242..2261
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2328..2430)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    2543..2576
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      2629..2985
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
       61 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
      121 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
      181 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
      241 cctctagagt cggtgggcct cgggggcggg tgcggggtcg gcggggccgc cccgggtggc
      301 ttcggtcgga gccatggggt cgtgcgctcc tttcggtcgg gcgctgcggg tcgtggggcg
      361 ggcgtcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga
      421 cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg
      481 tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg
      541 cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg
      601 cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc
      661 gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga
      721 cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc
      781 cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt
      841 ggcggtccgg gtcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg
      901 tgcgtacggc ccgggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg
      961 tcatggaagg tcgtctcctt gtgaggggtc aggggcgtgg gtcaggggat ggtggcggca
     1021 ccggtcgtgg cggccgacct gcaggcatgc aagctttttg caaaagccta ggcctccaaa
     1081 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     1141 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     1201 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     1261 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     1321 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     1381 acacacattc cacagatcaa gcttatcgat accgtcgacc tcgacctcca cggggagagc
     1441 ctgagcaaac tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     1501 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     1561 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     1621 gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     1681 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     1741 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     1801 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     1861 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     1921 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     1981 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     2041 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     2101 gccatacgta attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     2161 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     2221 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     2281 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     2341 gctcctgaaa atctcgacaa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     2401 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag
     2461 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc
     2521 actcgagccc ggggaattcc agataacttc gtataatgta tgctatacga agttatggta
     2581 ccgcggccgc gtagaggatc tgttgatcag cagttcaacc tgttgatagt acgtactaag
     2641 ctctcatgtt tcacgtacta agctctcatg tttaacgtac taagctctca tgtttaacga
     2701 actaaaccct catggctaac gtactaagct ctcatggcta acgtactaag ctctcatgtt
     2761 tcacgtacta agctctcatg tttgaacaat aaaattaata taaatcagca acttaaatag
     2821 cctctaaggt tttaagtttt ataagaaaaa aaagaatata taaggctttt aaagctttta
     2881 aggtttaacg gttgtggaca acaagccagg gatgtaacgc actgagaagc ccttagagcc
     2941 tctcaaagca attttcagtg acacaggaac acttaacggc tgacagaatt agcttcacgc
     3001 tgccgcaagc actcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt
     3061 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
     3121 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
     3181 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     3241 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
     3301 aggggatcaa
//
