LOCUS       sequence-260071-        7456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose inducible expression of lamba-Red recombinase proteins and
            I-SceI. Contains ampicillin resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7456)
  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 7456)
  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: pSNA
FEATURES             Location/Qualifiers
     source          1..7456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     256..275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(494..598)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(790..1668)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1695..1979
                     /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     1902..1921
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2768..3184
                     /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             3190..3975
                     /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             3972..4652
                     /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(5150..6100)
                     /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      6148..6370
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(join(7089..7456,1..424,425..493))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
       61 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
      121 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
      181 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
      241 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
      301 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
      361 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
      421 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
      481 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
      541 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
      601 catttccccg aaaagtgtcg tcggttcagg gcagggtcgt taaatagccg cttatgtcta
      661 ttgctggtct cggtacccgg ggatcctcta gagtcgacct gcataatgtg cctgtcaaat
      721 ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt
      781 cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg
      841 gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc
      901 gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag
      961 cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg
     1021 gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat
     1081 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
     1141 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
     1201 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
     1261 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
     1321 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
     1381 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
     1441 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
     1501 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
     1561 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
     1621 attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact
     1681 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
     1741 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
     1801 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
     1861 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
     1921 cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata
     1981 cccgtttttt tgggctagca ggagggtacc tatatgcata tgaaaatcat caaaaaaaac
     2041 caggtaatga acctcggtcc gaactctaaa ctgctgaaag aatacaaatc ccagctgatc
     2101 gaactgaaca tcgaacagtt cgaagcaggt atcggtctga tcctgggtga tgcttacatc
     2161 cgttctcgtg atgaaggtaa aacctactgt atgcagttcg agtggaaaaa caaagcatac
     2221 atggaccacg tatgtctgct gtacgatcag tgggtactgt ccccgccgca caaaaaagaa
     2281 cgtgttaacc acctgggtaa cctggtaatc acctggggcg cccagacttt caaacaccaa
     2341 gcattcaaca aactggctaa cctgttcatc gttaacaaca aaaaaaccat cccgaacaac
     2401 ctggttgaaa actacctgac cccgatgtct ctggcatact ggttcatgga tgatggtggt
     2461 aaatgggatt acaacaaaaa ctctaccaac aaatcgatcg tactgaacac ccagtctttc
     2521 actttcgaag aagtagaata cctggttaag ggtctgcgta acaaattcca actgaactgt
     2581 tacgtaaaaa tcaacaaaaa caaaccgatc atctacatcg attctatgtc ttacctgatc
     2641 ttctacaacc tgatcaaacc gtacctgatc ccgcagatga tgtacaaact gccgaacact
     2701 atctcctccg aaactttcct gaaataagct agaagatgtt tcgtgaagcc gtcgacgctt
     2761 ataaaaaatg gatattaata ctgaaactga gatcaagcaa aagcattcac taaccccctt
     2821 tcctgttttc ctaatcagcc cggcatttcg cgggcgatat tttcacagct atttcaggag
     2881 ttcagccatg aacgcttatt acattcagga tcgtcttgag gctcagagct gggcgcgtca
     2941 ctaccagcag ctcgcccgtg aagagaaaga ggcagaactg gcagacgaca tggaaaaagg
     3001 cctgccccag cacctgtttg aatcgctatg catcgatcat ttgcaacgcc acggggccag
     3061 caaaaaatcc attacccgtg cgtttgatga cgatgttgag tttcaggagc gcatggcaga
     3121 acacatccgg tacatggttg aaaccattgc tcaccaccag gttgatattg attcagaggt
     3181 ataaaacgaa tgagtactgc actcgcaacg ctggctggga agctggctga acgtgtcggc
     3241 atggattctg tcgacccaca ggaactgatc accactcttc gccagacggc atttaaaggt
     3301 gatgccagcg atgcgcagtt catcgcatta ctgatcgttg ccaaccagta cggccttaat
     3361 ccgtggacga aagaaattta cgcctttcct gataagcaga atggcatcgt tccggtggtg
     3421 ggcgttgatg gctggtcccg catcatcaat gaaaaccagc agtttgatgg catggacttt
     3481 gagcaggaca atgaatcctg tacatgccgg atttaccgca aggaccgtaa tcatccgatc
     3541 tgcgttaccg aatggatgga tgaatgccgc cgcgaaccat tcaaaactcg cgaaggcaga
     3601 gaaatcacgg ggccgtggca gtcgcatccc aaacggatgt tacgtcataa agccatgatt
     3661 cagtgtgccc gtctggcctt cggatttgct ggtatctatg acaaggatga agccgagcgc
     3721 attgtcgaaa atactgcata cactgcagaa cgtcagccgg aacgcgacat cactccggtt
     3781 aacgatgaaa ccatgcagga gattaacact ctgctgatcg ccctggataa aacatgggat
     3841 gacgacttat tgccgctctg ttcccagata tttcgccgcg acattcgtgc atcgtcagaa
     3901 ctgacacagg ccgaagcagt aaaagctctt ggattcctga aacagaaagc cgcagagcag
     3961 aaggtggcag catgacaccg gacattatcc tgcagcgtac cgggatcgat gtgagagctg
     4021 tcgaacaggg ggaagatgcg tggcacaaat tacggctcgg cgtcatcacc gcttcagaag
     4081 ttcacaacgt gatagcaaaa ccccgctccg gaaagaagtg gcctgacatg aaaatgtcct
     4141 acttccacac cctgcttgct gaggtttgca ccggtgtggc tccggaagtt aacgctaaag
     4201 cactggcctg gggaaaacag tacgagaacg acgccagaac cctgtttgaa ttcacttccg
     4261 gcgtgaatgt tactgaatcc ccgatcatct atcgcgacga aagtatgcgt accgcctgct
     4321 ctcccgatgg tttatgcagt gacggcaacg gccttgaact gaaatgcccg tttacctccc
     4381 gggatttcat gaagttccgg ctcggtggtt tcgaggccat aaagtcagct tacatggccc
     4441 aggtgcagta cagcatgtgg gtgacgcgaa aaaatgcctg gtactttgcc aactatgacc
     4501 cgcgtatgaa gcgtgaaggc ctgcattatg tcgtgattga gcgggatgaa aagtacatgg
     4561 cgagttttga cgagatcgtg ccggagttca tcgaaaaaat ggacgaggca ctggctgaaa
     4621 ttggttttgt atttggggag caatggcgat gacgcatcct cacgataata agcttcctgc
     4681 tgaacatcaa aggcaagaaa acatctgttg tcaaagacag catccttgaa caaggacaat
     4741 taacagttaa caaataaaaa cgcaaaagaa aatgccgata ttgactaccg gaagcagtgt
     4801 gaccgtgtgc ttctcaaatg cctgattcag gctgtctatg tgtgactgtt gagctgtaac
     4861 aagttgtctc aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt
     4921 tctattaggt gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagc
     4981 tttaaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc
     5041 atctgtgcat atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt
     5101 gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc
     5161 cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg
     5221 aaccattgag atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga
     5281 gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg
     5341 aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa
     5401 gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg
     5461 ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta
     5521 ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta
     5581 acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg
     5641 ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa
     5701 catgttccag attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt
     5761 cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa
     5821 tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg
     5881 ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc tgaacgtatt
     5941 ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga
     6001 gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa
     6061 tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt
     6121 gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc
     6181 tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg
     6241 ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg
     6301 ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg
     6361 tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt
     6421 ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct
     6481 cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt
     6541 tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac
     6601 tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc
     6661 acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct
     6721 gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag
     6781 gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt
     6841 cttactgtcg gggatcgacg ctctccctta tgcgactcct gcacctttcg tcttcgaata
     6901 aatacctgtg acggaagatc acttcgcaga ataaataaat cctggtctga cgctcagtgg
     6961 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7021 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7081 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7141 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7201 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7261 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7321 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7381 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     7441 gcttcattca gctccg
//
