LOCUS       sequence-14379-e        3546 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 3546)
  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 3546)
  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: pT-FLAG
FEATURES             Location/Qualifiers
     source          1..3546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             12..35
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(126..145)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(127..145)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    419..540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(456..475)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     510..529
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(547..1002)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(684..705)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     896..915
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1029..1133
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    1148..1181
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1223..2014,2015..2083))
                     /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     1846..1865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2253..2841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2742..2761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3016..3319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3320..3523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3474..3494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ctagcaccat ggattacaaa gacgatgacg ataaacctag cttcgaattc tgcagtcgac
       61 ggtaccgcgg gcccgggatc caccggatct agataatagg gtaccaggta agtgtaccca
      121 attcgcccta tagtgagtcg tattacaatt cactcgatcg cccttcccaa cagttgcgca
      181 gcctgaatgg cgaatggaga tccaattttt aagtgtataa tgtgttaaac tactgattct
      241 aattgtttgt gtattttaga ttcacagtcc caaggctcat ttcaggcccc tcagtcctca
      301 cagtctgttc atgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
      361 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
      421 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
      481 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
      541 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
      601 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      661 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      721 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      781 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      841 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      901 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      961 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     1021 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1081 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag aatcctgagg
     1141 ccgggccata acttcgtata atgtatgcta tacgaagtta tccatgggcc ccccctcgac
     1201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     1561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     1621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     1681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     1801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     1861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     1921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     1981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctcaggttac
     2101 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2161 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2221 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2281 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2341 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2401 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2461 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2521 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2581 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2641 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2701 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2761 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2821 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2881 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2941 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
     3001 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     3061 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     3121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     3181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     3241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     3301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     3361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     3421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     3481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     3541 gatccg
//
