LOCUS       sequence-288121-        7454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose inducible expression of lamba-Red recombinase proteins and
            I-SceI. Contains kamamycin resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7454)
  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 7454)
  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: pSNK
FEATURES             Location/Qualifiers
     source          1..7454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             430..846
                     /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             852..1637
                     /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"
     CDS             1634..2314
                     /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(2812..3762)
                     /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      3810..4032
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(4815..4834)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4826..5641
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4826..5641
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4899..4918)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(5906..6784)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        6811..7095
                     /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     7018..7037
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 aagcattcaa caaactggct aacctgttca tcgttaacaa caaaaaaacc atcccgaaca
       61 acctggttga aaactacctg accccgatgt ctctggcata ctggttcatg gatgatggtg
      121 gtaaatggga ttacaacaaa aactctacca acaaatcgat cgtactgaac acccagtctt
      181 tcactttcga agaagtagaa tacctggtta agggtctgcg taacaaattc caactgaact
      241 gttacgtaaa aatcaacaaa aacaaaccga tcatctacat cgattctatg tcttacctga
      301 tcttctacaa cctgatcaaa ccgtacctga tcccgcagat gatgtacaaa ctgccgaaca
      361 ctatctcctc cgaaactttc ctgaaataag ctagaagatg tttcgtgaag ccgtcgacgc
      421 ttataaaaaa tggatattaa tactgaaact gagatcaagc aaaagcattc actaaccccc
      481 tttcctgttt tcctaatcag cccggcattt cgcgggcgat attttcacag ctatttcagg
      541 agttcagcca tgaacgctta ttacattcag gatcgtcttg aggctcagag ctgggcgcgt
      601 cactaccagc agctcgcccg tgaagagaaa gaggcagaac tggcagacga catggaaaaa
      661 ggcctgcccc agcacctgtt tgaatcgcta tgcatcgatc atttgcaacg ccacggggcc
      721 agcaaaaaat ccattacccg tgcgtttgat gacgatgttg agtttcagga gcgcatggca
      781 gaacacatcc ggtacatggt tgaaaccatt gctcaccacc aggttgatat tgattcagag
      841 gtataaaacg aatgagtact gcactcgcaa cgctggctgg gaagctggct gaacgtgtcg
      901 gcatggattc tgtcgaccca caggaactga tcaccactct tcgccagacg gcatttaaag
      961 gtgatgccag cgatgcgcag ttcatcgcat tactgatcgt tgccaaccag tacggcctta
     1021 atccgtggac gaaagaaatt tacgcctttc ctgataagca gaatggcatc gttccggtgg
     1081 tgggcgttga tggctggtcc cgcatcatca atgaaaacca gcagtttgat ggcatggact
     1141 ttgagcagga caatgaatcc tgtacatgcc ggatttaccg caaggaccgt aatcatccga
     1201 tctgcgttac cgaatggatg gatgaatgcc gccgcgaacc attcaaaact cgcgaaggca
     1261 gagaaatcac ggggccgtgg cagtcgcatc ccaaacggat gttacgtcat aaagccatga
     1321 ttcagtgtgc ccgtctggcc ttcggatttg ctggtatcta tgacaaggat gaagccgagc
     1381 gcattgtcga aaatactgca tacactgcag aacgtcagcc ggaacgcgac atcactccgg
     1441 ttaacgatga aaccatgcag gagattaaca ctctgctgat cgccctggat aaaacatggg
     1501 atgacgactt attgccgctc tgttcccaga tatttcgccg cgacattcgt gcatcgtcag
     1561 aactgacaca ggccgaagca gtaaaagctc ttggattcct gaaacagaaa gccgcagagc
     1621 agaaggtggc agcatgacac cggacattat cctgcagcgt accgggatcg atgtgagagc
     1681 tgtcgaacag ggggaagatg cgtggcacaa attacggctc ggcgtcatca ccgcttcaga
     1741 agttcacaac gtgatagcaa aaccccgctc cggaaagaag tggcctgaca tgaaaatgtc
     1801 ctacttccac accctgcttg ctgaggtttg caccggtgtg gctccggaag ttaacgctaa
     1861 agcactggcc tggggaaaac agtacgagaa cgacgccaga accctgtttg aattcacttc
     1921 cggcgtgaat gttactgaat ccccgatcat ctatcgcgac gaaagtatgc gtaccgcctg
     1981 ctctcccgat ggtttatgca gtgacggcaa cggccttgaa ctgaaatgcc cgtttacctc
     2041 ccgggatttc atgaagttcc ggctcggtgg tttcgaggcc ataaagtcag cttacatggc
     2101 ccaggtgcag tacagcatgt gggtgacgcg aaaaaatgcc tggtactttg ccaactatga
     2161 cccgcgtatg aagcgtgaag gcctgcatta tgtcgtgatt gagcgggatg aaaagtacat
     2221 ggcgagtttt gacgagatcg tgccggagtt catcgaaaaa atggacgagg cactggctga
     2281 aattggtttt gtatttgggg agcaatggcg atgacgcatc ctcacgataa taagcttcct
     2341 gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac agcatccttg aacaaggaca
     2401 attaacagtt aacaaataaa aacgcaaaag aaaatgccga tattgactac cggaagcagt
     2461 gtgaccgtgt gcttctcaaa tgcctgattc aggctgtcta tgtgtgactg ttgagctgta
     2521 acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact ggtttcacct
     2581 gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc atggtgaaca
     2641 gctttaaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt
     2701 tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt
     2761 ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct
     2821 tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa
     2881 cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt
     2941 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta
     3001 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     3061 aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     3121 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     3181 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     3241 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     3301 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     3361 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     3421 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     3481 aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     3541 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgaacgta
     3601 ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt
     3661 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     3721 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     3781 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt
     3841 cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     3901 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     3961 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     4021 tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct
     4081 gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag
     4141 ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc
     4201 tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc
     4261 actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg
     4321 tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg
     4381 ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac
     4441 aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc
     4501 gtcttactgt cggggatcga cgctctccct tatgcgactc ctgcaccttt cgtcttcgaa
     4561 taaatacctg tgacggaaga tcacttcgca gataaaataa atcctggagt tggtagctct
     4621 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4681 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4741 cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac tgtctgctta
     4801 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag
     4861 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     4921 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
     4981 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     5041 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     5101 tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga
     5161 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     5221 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     5281 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     5341 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     5401 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     5461 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     5521 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     5581 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     5641 agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5701 ttccgcgcac atttccccga aaagtgccac ctgtcgtcgg ttcagggcag ggtcgttaaa
     5761 tagccgctta tgtctattgc tggtctcggt acccggggat cctctagagt cgacctgcat
     5821 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc
     5881 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
     5941 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
     6001 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
     6061 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
     6121 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
     6181 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
     6241 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
     6301 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
     6361 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
     6421 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
     6481 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
     6541 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
     6601 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
     6661 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
     6721 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     6781 ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat tgcatcagac
     6841 attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta accccgctta
     6901 ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt aacaaaagtg
     6961 tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca ctttgctatg
     7021 ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct
     7081 ctactgtttc tccatacccg tttttttggg ctagcaggag ggtacctata tgcatatgaa
     7141 aatcatcaaa aaaaaccagg taatgaacct cggtccgaac tctaaactgc tgaaagaata
     7201 caaatcccag ctgatcgaac tgaacatcga acagttcgaa gcaggtatcg gtctgatcct
     7261 gggtgatgct tacatccgtt ctcgtgatga aggtaaaacc tactgtatgc agttcgagtg
     7321 gaaaaacaaa gcatacatgg accacgtatg tctgctgtac gatcagtggg tactgtcccc
     7381 gccgcacaaa aaagaacgtg ttaaccacct gggtaacctg gtaatcacct ggggcgccca
     7441 gactttcaaa cacc
//
