LOCUS       sequence-296242-        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"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1074..1490
                     /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             1496..2281
                     /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             2278..2958
                     /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(3456..4406)
                     /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      4454..4676
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(join(5395..6186,6187..6255))
                     /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"
     primer_bind     6018..6037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6256..6360)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(6552..7430)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagcaggag ggtacctata tgcatatgaa aatcatcaaa aaaaaccagg taatgaacct
      361 cggtccgaac tctaaactgc tgaaagaata caaatcccag ctgatcgaac tgaacatcga
      421 acagttcgaa gcaggtatcg gtctgatcct gggtgatgct tacatccgtt ctcgtgatga
      481 aggtaaaacc tactgtatgc agttcgagtg gaaaaacaaa gcatacatgg accacgtatg
      541 tctgctgtac gatcagtggg tactgtcccc gccgcacaaa aaagaacgtg ttaaccacct
      601 gggtaacctg gtaatcacct ggggcgccca gactttcaaa caccaagcat tcaacaaact
      661 ggctaacctg ttcatcgtta acaacaaaaa aaccatcccg aacaacctgg ttgaaaacta
      721 cctgaccccg atgtctctgg catactggtt catggatgat ggtggtaaat gggattacaa
      781 caaaaactct accaacaaat cgatcgtact gaacacccag tctttcactt tcgaagaagt
      841 agaatacctg gttaagggtc tgcgtaacaa attccaactg aactgttacg taaaaatcaa
      901 caaaaacaaa ccgatcatct acatcgattc tatgtcttac ctgatcttct acaacctgat
      961 caaaccgtac ctgatcccgc agatgatgta caaactgccg aacactatct cctccgaaac
     1021 tttcctgaaa taagctagaa gatgtttcgt gaagccgtcg acgcttataa aaaatggata
     1081 ttaatactga aactgagatc aagcaaaagc attcactaac cccctttcct gttttcctaa
     1141 tcagcccggc atttcgcggg cgatattttc acagctattt caggagttca gccatgaacg
     1201 cttattacat tcaggatcgt cttgaggctc agagctgggc gcgtcactac cagcagctcg
     1261 cccgtgaaga gaaagaggca gaactggcag acgacatgga aaaaggcctg ccccagcacc
     1321 tgtttgaatc gctatgcatc gatcatttgc aacgccacgg ggccagcaaa aaatccatta
     1381 cccgtgcgtt tgatgacgat gttgagtttc aggagcgcat ggcagaacac atccggtaca
     1441 tggttgaaac cattgctcac caccaggttg atattgattc agaggtataa aacgaatgag
     1501 tactgcactc gcaacgctgg ctgggaagct ggctgaacgt gtcggcatgg attctgtcga
     1561 cccacaggaa ctgatcacca ctcttcgcca gacggcattt aaaggtgatg ccagcgatgc
     1621 gcagttcatc gcattactga tcgttgccaa ccagtacggc cttaatccgt ggacgaaaga
     1681 aatttacgcc tttcctgata agcagaatgg catcgttccg gtggtgggcg ttgatggctg
     1741 gtcccgcatc atcaatgaaa accagcagtt tgatggcatg gactttgagc aggacaatga
     1801 atcctgtaca tgccggattt accgcaagga ccgtaatcat ccgatctgcg ttaccgaatg
     1861 gatggatgaa tgccgccgcg aaccattcaa aactcgcgaa ggcagagaaa tcacggggcc
     1921 gtggcagtcg catcccaaac ggatgttacg tcataaagcc atgattcagt gtgcccgtct
     1981 ggccttcgga tttgctggta tctatgacaa ggatgaagcc gagcgcattg tcgaaaatac
     2041 tgcatacact gcagaacgtc agccggaacg cgacatcact ccggttaacg atgaaaccat
     2101 gcaggagatt aacactctgc tgatcgccct ggataaaaca tgggatgacg acttattgcc
     2161 gctctgttcc cagatatttc gccgcgacat tcgtgcatcg tcagaactga cacaggccga
     2221 agcagtaaaa gctcttggat tcctgaaaca gaaagccgca gagcagaagg tggcagcatg
     2281 acaccggaca ttatcctgca gcgtaccggg atcgatgtga gagctgtcga acagggggaa
     2341 gatgcgtggc acaaattacg gctcggcgtc atcaccgctt cagaagttca caacgtgata
     2401 gcaaaacccc gctccggaaa gaagtggcct gacatgaaaa tgtcctactt ccacaccctg
     2461 cttgctgagg tttgcaccgg tgtggctccg gaagttaacg ctaaagcact ggcctgggga
     2521 aaacagtacg agaacgacgc cagaaccctg tttgaattca cttccggcgt gaatgttact
     2581 gaatccccga tcatctatcg cgacgaaagt atgcgtaccg cctgctctcc cgatggttta
     2641 tgcagtgacg gcaacggcct tgaactgaaa tgcccgttta cctcccggga tttcatgaag
     2701 ttccggctcg gtggtttcga ggccataaag tcagcttaca tggcccaggt gcagtacagc
     2761 atgtgggtga cgcgaaaaaa tgcctggtac tttgccaact atgacccgcg tatgaagcgt
     2821 gaaggcctgc attatgtcgt gattgagcgg gatgaaaagt acatggcgag ttttgacgag
     2881 atcgtgccgg agttcatcga aaaaatggac gaggcactgg ctgaaattgg ttttgtattt
     2941 ggggagcaat ggcgatgacg catcctcacg ataataagct tcctgctgaa catcaaaggc
     3001 aagaaaacat ctgttgtcaa agacagcatc cttgaacaag gacaattaac agttaacaaa
     3061 taaaaacgca aaagaaaatg ccgatattga ctaccggaag cagtgtgacc gtgtgcttct
     3121 caaatgcctg attcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt
     3181 gttcaatttc atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta
     3241 catgctgttc atctgttaca ttgtcgatct gttcatggtg aacagcttta aatgcaccaa
     3301 aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg
     3361 acagttttcc ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg
     3421 atgcttcact gatagataca agagccataa gaacctcaga tccttccgta tttagccagt
     3481 atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca
     3541 tacttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca
     3601 gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg
     3661 gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga
     3721 tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta
     3781 tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc
     3841 cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat
     3901 tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat
     3961 aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta
     4021 tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat
     4081 tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta
     4141 accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat
     4201 acaccataag cattttccct actgatgttc atcatctgaa cgtattggtt ataagtgaac
     4261 gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag
     4321 cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt
     4381 gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta
     4441 taccaattga gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt
     4501 atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta
     4561 aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga
     4621 ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta
     4681 ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa
     4741 aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg
     4801 aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca
     4861 gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt
     4921 atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg
     4981 gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct
     5041 gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg
     5101 ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtcgggga
     5161 tcgacgctct cccttatgcg actcctgcac ctttcgtctt cgaataaata cctgtgacgg
     5221 aagatcactt cgcagaataa ataaatcctg gtctgacgct cagtggaacg aaaactcacg
     5281 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5341 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5401 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5461 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5521 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5581 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5641 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5701 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5761 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5821 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5881 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5941 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6001 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6061 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6121 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6181 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6241 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6301 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6361 cacatttccc cgaaaagtgt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc
     6421 tattgctggt ctcggtaccc ggggatcctc tagagtcgac ctgcataatg tgcctgtcaa
     6481 atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc aattgtctga
     6541 ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca
     6601 cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga
     6661 tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc
     6721 agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc
     6781 tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg
     6841 atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga
     6901 ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc
     6961 tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg
     7021 atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc
     7081 gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag
     7141 taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct
     7201 cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc
     7261 accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg
     7321 tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg
     7381 gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata
     7441 ctcccgccat tcagag
//
