LOCUS       sequence-7131-ex        3807 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 3807)
  AUTHORS   Sauer B
  TITLE     Manipulation of transgenes by site-specific recombination: use of 
            Cre recombinase.
  JOURNAL   Methods Enzymol. 1993;225:890-900. doi: 
            10.1016/0076-6879(93)25056-8.
  PUBMED    8231893
REFERENCE   2  (bases 1 to 3807)
  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: "Methods 
            Enzymol."; date: "1993"; volume: "225:890-900. doi"; pages: " 
            10.1016/0076-6879(93)25056-8"
COMMENT     Sequence Label: pBS302
FEATURES             Location/Qualifiers
     source          1..3807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     50..68
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    118..151
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    193..327
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(217..236)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     271..290
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(752..772)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          902..967
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     protein_bind    1501..1534
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1619..1641)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1727..1867
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1782..1801
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1882..1899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2053..2641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2133..2152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2812..3603,3604..3672))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3435..3454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3673..3777)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcggccgcac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
       61 cacgaggccc tttcgtcttc aagaattcca tcaagcttag gatccggaac ccttaatata
      121 acttcgtata atgtatgcta tacgaagtta ttaggtccct cgacctgcag cccaagctta
      181 cttaccatgt cagatccaga catgataaga tacattgatg agtttggaca aaccacaact
      241 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
      301 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
      361 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
      421 gctgattatg atctctagtc aaggcactat acatcaaata ttccttatta acccctttac
      481 aaattaaaaa gctaaaggta cacaattttt gagcatagtt attaatagca gacactctat
      541 gcctgtgtgg agtaagaaaa aacagtatgt tatgattata actgttatgc ctacttataa
      601 aggttacaga atatttttcc ataattttct tgtatagcag tgcagctttt tcctttgtgg
      661 tgtaaatagc aaagcaagca agagttctat tactaaacac agcatgactc aaaaaactta
      721 gcaattctga aggaaagtcc ttggggtctt ctacctttct cttctttttt ggaggagtag
      781 aatgttgaga gtcagcagta gcctcatcat cactagatgg catttcttct gagcaaaaca
      841 ggttttcctc attaaaggca ttccaccact gctcccattc atcagttcca taggttggaa
      901 tctaaaatac acaaacaatt agaatcagta gtttaacaca ttatacactt aaaaatttta
      961 tatttacctt agagctttaa atctctgtag gtagtttgtc caattatgtc acaccacaga
     1021 agtaaggttc cttcacaaag atccctcgag aaaaaaaata taaaagagat ggaggaacgg
     1081 gaaaaagtta gttgtggtga taggtggcaa gtggtattcc gtaagaacaa caagaaaagc
     1141 atttcatatt atggctgaac tgagcgaaca agtgcaaaat ttaagcatca acgacaacaa
     1201 cgagaatggt tatgttcctc ctcacttaag aggaaaacca agaagtgcca gaaataacat
     1261 gagcaactac aataacaaca acggcggcta caacggtggc cgtggcggtg gcagcttctt
     1321 tagcaacaac cgtcgtggtg gttacggcaa cggtggtttc ttcggtggaa acaacggtgg
     1381 cagcagatct aacggccgtt ctggtggtag atggatcgat ggcaaacatg tcccagctcc
     1441 aagaaacgaa aaggccgaga tcgccatatt tggtgtcccc gaggatccgg aacccttaat
     1501 ataacttcgt ataatgtatg ctatacgaag ttattaggtc cctcgaagag gttcactagt
     1561 actggccatt gcggccgctt cgaggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     1621 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     1681 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     1741 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     1801 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     1861 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2221 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     2281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2461 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2701 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2761 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2821 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2881 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2941 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3001 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3061 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3121 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3181 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3241 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3301 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3361 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3421 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3481 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3541 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3601 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3661 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3781 atttccccga aaagtgccac ctgacgt
//
