LOCUS       sequence-338839-        7288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo Cre-lox circularization in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7288)
  AUTHORS   Enghiad B, Huang C, Guo F, Jiang G, Wang B, Tabatabaei SK, Martin 
            TA, Zhao H
  TITLE     Cas12a-assisted precise targeted cloning using in vivo Cre-lox 
            recombination.
  JOURNAL   Nat Commun. 2021 Feb 19;12(1):1171. doi: 10.1038/s41467-021-21275-4.
  PUBMED    33608525
REFERENCE   2  (bases 1 to 7288)
  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: "Nat 
            Commun."; date: "2021-02-19"; volume: "12(1):1171. doi"; pages: " 
            10.1038/s41467-021-21275-4"
COMMENT     Sequence Label: pBE14
FEATURES             Location/Qualifiers
     source          1..7288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(11..889)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        916..1200
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1123..1142
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1227..1235
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1244..1660
                     /codon_start=1
                     /gene="gam"
                     /product="inhibitor of the host RecBCD nuclease in the 
                     lambda Red system"
                     /label=Gam
                     /note="protects the double-stranded DNA substrate for 
                     recombination"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     RBS             1661..1672
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1673..2734
                     /codon_start=1
                     /gene="recA"
                     /product="DNA repair protein from E. coli"
                     /label=RecA
                     /translation="MAIDENKQKALAAALGQIEKQFGKGSIMRLGEDRSMDVETISTGS
                     LSLDIALGAGGLPMGRIVEIYGPESSGKTTLTLQVIAAAQREGKTCAFIDAEHALDPIY
                     ARKLGVDIDNLLCSQPDTGEQALEICDALARSGAVDVIVVDSVAALTPKAEIEGEIDDS
                     HMGLAARMMSQAMRKLAGNLKQSNTLLIFINQIRMKIGVMFGNPETTTGGNALKFYASV
                     RLDIRRIGAVKEGENVVGSETRVKVVKNKIAAPFKQAEFQILYGEGINFYGELVDLGVK
                     EKLIEKAGAWYSYKGEKIGQGKANATAWLKDNPETAKEIEKKVRELLLSNPNSTPDFSV
                     DDSEGVAETNEDF"
     terminator      2735..2979
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(3102..4052)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4100..4322
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        4576..4610
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             4611..4622
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4623..5654
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     terminator      5655..5689
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(5988..7178)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     6807..6826
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(7226..7231,7232..7248,7249..7254))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 catcgattta ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
       61 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
      121 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
      181 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
      241 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
      301 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
      361 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
      421 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
      481 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
      541 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
      601 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
      661 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
      721 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
      781 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
      841 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
      901 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
      961 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
     1021 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
     1081 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
     1141 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
     1201 acccgttttt ttgggaattc gagctctaag gaggttataa aaaatggata ttaatactga
     1261 aactgagatc aagcaaaagc attcactaac cccctttcct gttttcctaa tcagcccggc
     1321 atttcgcggg cgatattttc acagctattt caggagttca gccatgaacg cttattacat
     1381 tcaggatcgt cttgaggctc agagctgggc gcgtcactac cagcagctcg cccgtgaaga
     1441 gaaagaggca gaactggcag acgacatgga aaaaggcctg ccccagcacc tgtttgaatc
     1501 gctatgcatc gatcatttgc aacgccacgg ggccagcaaa aaatccatta cccgtgcgtt
     1561 tgatgacgat gttgagtttc aggagcgcat ggcagaacac atccggtaca tggttgaaac
     1621 cattgctcac caccaggttg atattgattc agaggtataa aaagaggaga aaatggctat
     1681 cgacgaaaac aaacagaaag cgttggcggc agcactgggc cagattgaga aacaatttgg
     1741 taaaggctcc atcatgcgcc tgggtgaaga ccgttccatg gatgtggaaa ccatctctac
     1801 cggttcgctt tcactggata tcgcgcttgg ggcaggtggt ctgccgatgg gccgtatcgt
     1861 cgaaatctac ggaccggaat cttccggtaa aaccacgctg acgctgcagg tgatcgccgc
     1921 agcgcagcgt gaaggtaaaa cctgtgcgtt tatcgatgct gaacacgcgc tggacccaat
     1981 ctacgcacgt aaactgggcg tcgatatcga caacctgctg tgctcccagc cggacaccgg
     2041 cgagcaggca ctggaaatct gtgacgccct ggcgcgttct ggcgcagtag acgttatcgt
     2101 cgttgactcc gtggcggcac tgacgccgaa agcggaaatc gaaggcgaaa tcgacgactc
     2161 tcacatgggc cttgcggcac gtatgatgag ccaggcgatg cgtaagctgg cgggtaacct
     2221 gaagcagtcc aacacgctgc tgatcttcat caaccagatc cgtatgaaaa ttggtgtgat
     2281 gttcggtaac ccggaaacca ctaccggtgg taacgcgctg aaattctacg cctctgttcg
     2341 tctcgacatc cgtcgtatcg gcgcggtgaa agagggcgaa aacgtggtgg gtagcgaaac
     2401 ccgcgtgaaa gtggtgaaga acaaaatcgc tgcgccgttt aaacaggctg aattccagat
     2461 cctctacggc gaaggtatca acttctacgg cgaactggtt gacctgggcg taaaagagaa
     2521 gctgatcgag aaagcaggcg cgtggtacag ctacaaaggt gagaagatcg gtcagggtaa
     2581 agcgaatgcg actgcctggc tgaaagataa cccggaaacc gcgaaagaga tcgagaagaa
     2641 agtacgtgag ttgctgctga gcaacccgaa ctcaacgccg gatttctctg tagatgatag
     2701 cgaaggcgta gcagaaacta acgaagattt ttaacgcatc ctcacgataa tatccgggta
     2761 ggcgcaatca ctttcgtcta ctccgttaca aagcgaggct gggtatttcc cggcctttct
     2821 gttatccgaa atccactgaa agcacagcgg ctggctgagg agataaataa taaacgaggg
     2881 gctgtatgca caaagcatct tctgttgagt taagaacgag tatcgagatg gcacatagcc
     2941 ttgctcaaat tggaatcagg tttgtgccaa taccagtaga aacagacgaa gaatccatgg
     3001 gtatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt ttgtttgtta
     3061 gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta
     3121 gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg
     3181 agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt
     3241 tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tacgtaggta ggaatctgat
     3301 gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt
     3361 acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct
     3421 cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact tattggtttc
     3481 aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac
     3541 ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct
     3601 tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc
     3661 agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa
     3721 actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag
     3781 cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta
     3841 gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa
     3901 gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc
     3961 acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt
     4021 tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa
     4081 tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt cctttgagtt
     4141 gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc
     4201 tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt
     4261 tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac
     4321 tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct
     4381 ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcggttgcg
     4441 gccgcaatcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc
     4501 gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta
     4561 aatggcacta caggcttgac agctagctca gtcctaggta taatgctagc aaagaggaga
     4621 aaatgtccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc gatgcaacga
     4681 gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg ttttctgagc
     4741 atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc aagttgaata
     4801 accggaaatg gtttcccgca gaacctgaag atgttcgcga ttatcttcta tatcttcagg
     4861 cgcgcggtct ggcagtaaaa actatccagc aacatttggg ccagctaaac atgcttcatc
     4921 gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt ttcactggtt atgcggcgga
     4981 tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg ttcgaacgca
     5041 ctgatttcga ccaggttcgt tcactcatgg aaaatagcga tcgctgccag gatatacgta
     5101 atctggcatt tctggggatt gcttataaca ccctgttacg tatagccgaa attgccagga
     5161 tcagggttaa agatatctca cgtactgacg gtgggagaat gttaatccat attggcagaa
     5221 cgaaaacgct ggttagcacc gcaggtgtag agaaggcact tagcctgggg gtaactaaac
     5281 tggtcgagcg atggatttcc gtctctggtg tagctgatga tccgaataac tacctgtttt
     5341 gccgggtcag aaaaaatggt gttgccgcgc catctgccac cagccagcta tcaactcgcg
     5401 ccctggaagg gatttttgaa gcaactcatc gattgattta cggcgctaag gatgactctg
     5461 gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg cgagatatgg
     5521 cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc aatgtaaata
     5581 ttgtcatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg cgcctgctgg
     5641 aagatggcga ttagttcaga acgctcggtt gccgccgggc gttttttatg ccttttatgg
     5701 attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt
     5761 tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc
     5821 tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa
     5881 tgcacccagt aaggcagcgg tatcatcaac ggggtctgac gctcagtgga acgaaaactc
     5941 acgtcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg
     6001 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct
     6061 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc
     6121 agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc
     6181 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
     6241 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc
     6301 aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     6361 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     6421 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     6481 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     6541 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     6601 ggcatcggtc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     6661 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     6721 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     6781 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     6841 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcca caggacgggt
     6901 gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag caggactggg
     6961 cggcggccaa agcggtcgga cagtgctccg agaacgggtg cgcatagaaa ttgcatcaac
     7021 gcatatagcg ctagcagcac gccatagtga ctggcgatgc tgtcggaatg gacgatatcc
     7081 cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg
     7141 gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag
     7201 caatttaact gtgataaact accgcattaa agcttatcga tgataagctg tcaaacatga
     7261 gacacatttc cccgaaaagt gccacctg
//
