LOCUS       sequence-14376-e        5325 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 5325)
  AUTHORS   Werdien D, Peiler G, Ryffel GU
  TITLE     FLP and Cre recombinase function in Xenopus embryos.
  JOURNAL   Nucleic Acids Res. 2001 Jun 1. 29(11):E53-3.
  PUBMED    11376165
REFERENCE   2  (bases 1 to 5325)
  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: "Nucleic 
            Acids Res. 2001 Jun 1. 29(11):E53-3."
COMMENT     Sequence Label: LCMV:ECFP(loxP)MCS
FEATURES             Location/Qualifiers
     source          1..5325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(34..67)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      78..87
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(84..86,87..89,90..803)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(129..150)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(390..409)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     737..758
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    917..1051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1053..1086)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          1207..1272
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    1319..1453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1607..1627)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1607..1625)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1646..1662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1646..1662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1659..1681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1670..1686
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1694..1700,1701..1718,1719..1724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1739..1760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1877..1894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2048..2636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2128..2147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2807..3598,3599..3667))
                     /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     3430..3449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3668..3772)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3798..4253)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3935..3956)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4147..4166
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4341..5325
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttataactt cgtatagcat acattatacg
       61 aagttatagg atccatcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      121 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      181 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      241 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca
      301 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      361 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      421 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      481 aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata
      541 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
      601 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
      661 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
      721 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
      781 gcatggacga gctgtacaag taatctagag gatcataatc agccatacca catttgtaga
      841 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
      901 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
      961 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1021 actcatcaat gtatcttatc atgtctggat ccataacttc gtatagcata cattatacga
     1081 agttatacct aggagatctc accggtgact agtcaggggg gatctgcatg gatctttgtg
     1141 aaggaacctt acttctgtgg tgtgacataa ttggacaaac tacctacaga gatttaaagc
     1201 tctaaggtaa atataaaatt tttaagtgta taatgtgtta aactactgat tctaattgtt
     1261 tgtgtatttt agattccaac ctatggaact gatgaatggg agcagtggtg gaatgccaga
     1321 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1381 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1441 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1501 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatct
     1561 gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt
     1621 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     1681 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     1741 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     1801 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     1861 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     1921 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     1981 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2041 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2101 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2161 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2221 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2281 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2341 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2401 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2461 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     2521 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2581 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2641 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2701 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2761 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2821 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2881 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     2941 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3001 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3061 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3121 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3181 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3241 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3301 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3361 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3421 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3481 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3541 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3601 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3661 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3721 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     3781 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3841 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3901 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3961 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4021 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4081 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4141 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4201 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     4261 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     4321 ccagtcgagg cggccgcgtc gaccatagcc aattcaatat ggcgtatatg gactcatgcc
     4381 aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc
     4441 aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc
     4501 aattcaatat ggcggacttg gcactgtgcc aactggggag gggtctactt ggcacggtgc
     4561 caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc
     4621 caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg
     4681 gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat
     4741 agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca
     4801 tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg
     4861 tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg
     4921 cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc
     4981 aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct
     5041 cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta
     5101 ttaatagtaa cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac
     5161 tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga
     5221 cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg
     5281 cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaac
//
