LOCUS       sequence-273788-        7312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose inducible expression of lamba-Red recombinase proteins and
            I-CreI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7312)
  AUTHORS   Wei Y, Deng P, Mohsin A, Yang Y, Zhou H, Guo M, Fang H
  TITLE     An electroporation-free method based on Red recombineering for 
            markerless deletion and genomic replacement in the Escherichia coli 
            DH1 genome.
  JOURNAL   PLoS One. 2017 Oct 24;12(10):e0186891. doi: 
            10.1371/journal.pone.0186891. eCollection 2017.
  PUBMED    29065183
REFERENCE   2  (bases 1 to 7312)
  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: "PLoS One.";
            date: "2017-10-24"; volume: "12(10):e0186891. doi"; pages: " 
            10.1371/journal.pone.0186891. eCollection 2017"
COMMENT     Sequence Label: pCNA
FEATURES             Location/Qualifiers
     source          1..7312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             473..1153
                     /codon_start=1
                     /gene="exo"
                     /product="5' to 3' double-stranded DNA exonuclease in the 
                     lambda Red system"
                     /label=Exo
                     /note="creates 3' single-stranded DNA tails"
                     /translation="MTPDIILQRTGIDVRAVEQGEDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     CDS             complement(1651..2601)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2649..2871
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(3464..3482)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3550..3654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3655..3723,3724..4515)
                     /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     complement(3873..3892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4835..5713)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5740..6024
                     /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     5947..5966
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             6581..6997
                     /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"
     CDS             join(7003..7312,1..476)
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
ORIGIN
        1 gtacatgccg gatttaccgc aaggaccgta atcatccgat ctgcgttacc gaatggatgg
       61 atgaatgccg ccgcgaacca ttcaaaactc gcgaaggcag agaaatcacg gggccgtggc
      121 agtcgcatcc caaacggatg ttacgtcata aagccatgat tcagtgtgcc cgtctggcct
      181 tcggatttgc tggtatctat gacaaggatg aagccgagcg cattgtcgaa aatactgcat
      241 acactgcaga acgtcagccg gaacgcgaca tcactccggt taacgatgaa accatgcagg
      301 agattaacac tctgctgatc gccctggata aaacatggga tgacgactta ttgccgctct
      361 gttcccagat atttcgccgc gacattcgtg catcgtcaga actgacacag gccgaagcag
      421 taaaagctct tggattcctg aaacagaaag ccgcagagca gaaggtggca gcatgacacc
      481 ggacattatc ctgcagcgta ccgggatcga tgtgagagct gtcgaacagg gggaagatgc
      541 gtggcacaaa ttacggctcg gcgtcatcac cgcttcagaa gttcacaacg tgatagcaaa
      601 accccgctcc ggaaagaagt ggcctgacat gaaaatgtcc tacttccaca ccctgcttgc
      661 tgaggtttgc accggtgtgg ctccggaagt taacgctaaa gcactggcct ggggaaaaca
      721 gtacgagaac gacgccagaa ccctgtttga attcacttcc ggcgtgaatg ttactgaatc
      781 cccgatcatc tatcgcgacg aaagtatgcg taccgcctgc tctcccgatg gtttatgcag
      841 tgacggcaac ggccttgaac tgaaatgccc gtttacctcc cgggatttca tgaagttccg
      901 gctcggtggt ttcgaggcca taaagtcagc ttacatggcc caggtgcagt acagcatgtg
      961 ggtgacgcga aaaaatgcct ggtactttgc caactatgac ccgcgtatga agcgtgaagg
     1021 cctgcattat gtcgtgattg agcgggatga aaagtacatg gcgagttttg acgagatcgt
     1081 gccggagttc atcgaaaaaa tggacgaggc actggctgaa attggttttg tatttgggga
     1141 gcaatggcga tgacgcatcc tcacgataat aagcttcctg ctgaacatca aaggcaagaa
     1201 aacatctgtt gtcaaagaca gcatccttga acaaggacaa ttaacagtta acaaataaaa
     1261 acgcaaaaga aaatgccgat attgactacc ggaagcagtg tgaccgtgtg cttctcaaat
     1321 gcctgattca ggctgtctat gtgtgactgt tgagctgtaa caagttgtct caggtgttca
     1381 atttcatgtt ctagttgctt tgttttactg gtttcacctg ttctattagg tgttacatgc
     1441 tgttcatctg ttacattgtc gatctgttca tggtgaacag ctttaaatgc accaaaaact
     1501 cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt
     1561 tttccctttg atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct
     1621 tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt
     1681 ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt
     1741 actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa
     1801 agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt
     1861 tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat
     1921 ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac
     1981 caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg
     2041 gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc
     2101 aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca
     2161 ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt
     2221 tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa
     2281 tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa
     2341 aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc
     2401 ataagcattt tccctactga tgttcatcat ctgaacgtat tggttataag tgaacgatac
     2461 cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa
     2521 aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt
     2581 gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca
     2641 attgagatgg gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg
     2701 taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc
     2761 cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa
     2821 gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta
     2881 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
     2941 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat
     3001 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     3061 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga
     3121 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     3181 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     3241 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat
     3301 gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc ggggatcgac
     3361 gctctccctt atgcgactcc tgcacctttc gtcttcgaat aaatacctgt gacggaagat
     3421 cacttcgcag aataaataaa tcctggtgtc cctggacgaa agggcctcgt gatacgccta
     3481 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3541 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3601 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3661 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3721 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3781 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3841 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3901 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3961 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4021 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4081 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4141 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4201 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4261 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4321 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4381 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4441 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4501 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgcgtaattt
     4561 ttttaaggca gttattggtg cccttaaacg cctggttgct acgcctgaat aagtgataat
     4621 aagcggatga atggcagaaa ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg
     4681 gtcgttaaat agccgcttat gtctattgct ggtctcggta cccggggatc ctctagagtc
     4741 gacctgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
     4801 ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat
     4861 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
     4921 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
     4981 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
     5041 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
     5101 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
     5161 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
     5221 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
     5281 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
     5341 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
     5401 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
     5461 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
     5521 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
     5581 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
     5641 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
     5701 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt
     5761 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa
     5821 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
     5881 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
     5941 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
     6001 cgcaactctc tactgtttct ccatacccgt ttttttgggc tagctaagaa ggagatatac
     6061 atatgttata cctggcaggt ttcgtggacg gtgacggtag catcatcgct cagattaaac
     6121 caaaccagtc ttataagttt aaacatcagc tgagcttgac ctttcaggtg actcaaaaga
     6181 cccagcgccg ttggtttctg gacaaactag tggatgaaat tggcgttggt tacgtacgtg
     6241 atcgcggttc cgtttccgat tacatcttaa gcgaaatcaa gccgctgcac aacttcctga
     6301 ctcaactgca gccgtttctg aaactgaaac agaaacaggc aaacctggtt ctgaaaatta
     6361 tcgaacagct gccgtctgca aaagaatccc cggacaaatt cctggaagtt tgtacctggg
     6421 tggatcagat tgcagctctg aacgattcta agactcgtaa aaccacttct gaaaccgttc
     6481 gtgctgtgct ggacagcctg agcgagaaga agaaatcctc cccggcggcc gactaatcta
     6541 gcgaaaagat gtttcgtgaa gccgtcgacg cttataaaaa atggatatta atactgaaac
     6601 tgagatcaag caaaagcatt cactaacccc ctttcctgtt ttcctaatca gcccggcatt
     6661 tcgcgggcga tattttcaca gctatttcag gagttcagcc atgaacgctt attacattca
     6721 ggatcgtctt gaggctcaga gctgggcgcg tcactaccag cagctcgccc gtgaagagaa
     6781 agaggcagaa ctggcagacg acatggaaaa aggcctgccc cagcacctgt ttgaatcgct
     6841 atgcatcgat catttgcaac gccacggggc cagcaaaaaa tccattaccc gtgcgtttga
     6901 tgacgatgtt gagtttcagg agcgcatggc agaacacatc cggtacatgg ttgaaaccat
     6961 tgctcaccac caggttgata ttgattcaga ggtataaaac gaatgagtac tgcactcgca
     7021 acgctggctg ggaagctggc tgaacgtgtc ggcatggatt ctgtcgaccc acaggaactg
     7081 atcaccactc ttcgccagac ggcatttaaa ggtgatgcca gcgatgcgca gttcatcgca
     7141 ttactgatcg ttgccaacca gtacggcctt aatccgtgga cgaaagaaat ttacgccttt
     7201 cctgataagc agaatggcat cgttccggtg gtgggcgttg atggctggtc ccgcatcatc
     7261 aatgaaaacc agcagtttga tggcatggac tttgagcagg acaatgaatc ct
//
