LOCUS       sequence-380750-        7754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the lambda Red recombinase under control of the 
            arabinose-inducible P(araB) promoter and I-SceI under control of the
            anhydrotetracycline-inducible P(tetA) promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7754)
  AUTHORS   Kim J, Webb AM, Kershner JP, Blaskowski S, Copley SD
  TITLE     A versatile and highly efficient method for scarless genome editing 
            in Escherichia coli and Salmonella enterica.
  JOURNAL   BMC Biotechnol. 2014 Sep 25;14(1):84.
  PUBMED    25255806
REFERENCE   2  (bases 1 to 7754)
  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: "BMC 
            Biotechnol."; date: "2014-09-25"; volume: "14(1)"; pages: "84"
COMMENT     Sequence Label: pSLTS
FEATURES             Location/Qualifiers
     source          1..7754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     493..512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(731..835)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(872..1750)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1777..2061
                     /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     1984..2003
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             2088..2096
                     /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             2105..2521
                     /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             2527..3312
                     /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             3309..3989
                     /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="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      3990..4234
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(4256..4882)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     4787..4806
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        4898..4953
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    4904..4922
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    4934..4952
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(5782..6732)
                     /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      6780..7002
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(join(7624..7754,1..661,662..730))
                     /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"
ORIGIN
        1 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
       61 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
      121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
      181 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
      241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
      301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
      361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
      421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
      481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
      541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
      601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
      661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
      721 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      841 ttccccgaaa agtgccacct gcatcgattt attatgacaa cttgacggct acatcattca
      901 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
      961 cccgcgagaa gtagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     1021 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     1081 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     1141 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     1201 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     1261 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     1321 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     1381 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     1441 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     1501 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     1561 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     1621 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     1681 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     1741 cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca
     1801 tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc
     1861 cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca
     1921 aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt
     1981 gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc
     2041 aactctctac tgtttctcca tacccgtttt tttgggaatt cgagctctaa ggaggttata
     2101 aaaaatggat attaatactg aaactgagat caagcaaaag cattcactaa ccccctttcc
     2161 tgttttccta atcagcccgg catttcgcgg gcgatatttt cacagctatt tcaggagttc
     2221 agccatgaac gcttattaca ttcaggatcg tcttgaggct cagagctggg cgcgtcacta
     2281 ccagcagctc gcccgtgaag agaaagaggc agaactggca gacgacatgg aaaaaggcct
     2341 gccccagcac ctgtttgaat cgctatgcat cgatcatttg caacgccacg gggccagcaa
     2401 aaaatccatt acccgtgcgt ttgatgacga tgttgagttt caggagcgca tggcagaaca
     2461 catccggtac atggttgaaa ccattgctca ccaccaggtt gatattgatt cagaggtata
     2521 aaacgaatga gtactgcact cgcaacgctg gctgggaagc tggctgaacg tgtcggcatg
     2581 gattctgtcg acccacagga actgatcacc actcttcgcc agacggcatt taaaggtgat
     2641 gccagcgatg cgcagttcat cgcattactg atcgttgcca accagtacgg ccttaatccg
     2701 tggacgaaag aaatttacgc ctttcctgat aagcagaatg gcatcgttcc ggtggtgggc
     2761 gttgatggct ggtcccgcat catcaatgaa aaccagcagt ttgatggcat ggactttgag
     2821 caggacaatg aatcctgtac atgccggatt taccgcaagg accgtaatca tccgatctgc
     2881 gttaccgaat ggatggatga atgccgccgc gaaccattca aaactcgcga aggcagagaa
     2941 atcacggggc cgtggcagtc gcatcccaaa cggatgttac gtcataaagc catgattcag
     3001 tgtgcccgtc tggccttcgg atttgctggt atctatgaca aggatgaagc cgagcgcatt
     3061 gtcgaaaata ctgcatacac tgcagaacgt cagccggaac gcgacatcac tccggttaac
     3121 gatgaaacca tgcaggagat taacactctg ctgatcgccc tggataaaac atgggatgac
     3181 gacttattgc cgctctgttc ccagatattt cgccgcgaca ttcgtgcatc gtcagaactg
     3241 acacaggccg aagcagtaaa agctcttgga ttcctgaaac agaaagccgc agagcagaag
     3301 gtggcagcat gacaccggac attatcctgc agcgtaccgg gatcgatgtg agagctgtcg
     3361 aacaggggga tgatgcgtgg cacaaattac ggctcggcgt catcaccgct tcagaagttc
     3421 acaacgtgat agcaaaaccc cgctccggaa agaagtggcc tgacatgaaa atgtcctact
     3481 tccacaccct gcttgctgag gtttgcaccg gtgtggctcc ggaagttaac gctaaagcac
     3541 tggcctgggg aaaacagtac gagaacgacg ccagaaccct gtttgaattc acttccggcg
     3601 tgaatgttac tgaatccccg atcatctatc gcgacgaaag tatgcgtacc gcctgctctc
     3661 ccgatggttt atgcagtgac ggcaacggcc ttgaactgaa atgcccgttt acctcccggg
     3721 atttcatgaa gttccggctc ggtggtttcg aggccataaa gtcagcttac atggcccagg
     3781 tgcagtacag catgtgggtg acgcgaaaaa atgcctggta ctttgccaac tatgacccgc
     3841 gtatgaagcg tgaaggcctg cattatgtcg tgattgagcg ggatgaaaag tacatggcga
     3901 gttttgacga gatcgtgccg gagttcatcg aaaaaatgga cgaggcactg gctgaaattg
     3961 gttttgtatt tggggagcaa tggcgatgac gcatcctcac gataatatcc gggtaggcgc
     4021 aatcactttc gtctactccg ttacaaagcg aggctgggta tttcccggcc tttctgttat
     4081 ccgaaatcca ctgaaagcac agcggctggc tgaggagata aataataaac gaggggctgt
     4141 atgcacaaag catcttctgt tgagttaaga acgagtatcg agatggcaca tagccttgct
     4201 caaattggaa tcaggtttgt gccaatacca gtagaaacag acgaagaatc catggttaag
     4261 acccactttc acatttaagt tgtttttcta atccgcatat gatcaattca aggccgaata
     4321 agaaggctgg ctctgcacct tggtgatcaa ataattcgat agcttgtcgt aataatggcg
     4381 gcatactatc agtagtaggt gtttcccttt cttctttagc gacttgatgc tcttgatctt
     4441 ccaatacgca acctaaagta aaatgcccca cagcgctgag tgcatataat gcattctcta
     4501 gtgaaaaacc ttgttggcat aaaaaggcta attgattttc gagagtttca tactgttttt
     4561 ctgtaggccg tgtacctaaa tgtacttttg ctccatcgcg atgacttagt aaagcacatc
     4621 taaaactttt agcgttatta cgtaaaaaat cttgccagct ttccccttct aaagggcaaa
     4681 agtgagtatg gtgcctatct aacatctcaa tggctaaggc gtcgagcaaa gcccgcttat
     4741 tttttacatg ccaatacaat gtaggctgct ctacacctag cttctgggcg agtttacggg
     4801 ttgttaaacc ttcgattccg acctcattaa gcagctctaa tgcgctgtta atcactttac
     4861 ttttatctaa tctagacatc attaattcct aatttttgtt gacactctat cattgataga
     4921 gttattttac cactccctat cagtgataga gaaaagtgaa atgcatatga aaaacatcaa
     4981 aaaaaaccag gtaatgaacc tgggtccgaa ctctaaactg ctgaaagaat acaaatccca
     5041 gctgatcgaa ctgaacatcg aacagttcga agcaggtatc ggtctgatcc tgggtgatgc
     5101 ttacatccgt tctcgtgatg aaggtaaaac ctactgtatg cagttcgagt ggaaaaacaa
     5161 agcatacatg gaccacgtat gtctgctgta cgatcagtgg gtactgtccc cgccgcacaa
     5221 aaaagaacgt gttaaccacc tgggtaacct ggtaatcacc tggggcgccc agactttcaa
     5281 acaccaagct ttcaacaaac tggctaacct gttcatcgtt aacaacaaaa aaaccatccc
     5341 gaacaacctg gttgaaaact acctgacccc gatgtctctg gcatactggt tcatggatga
     5401 tggtggtaaa tgggattaca acaaaaactc taccaacaaa tcgatcgtac tgaacaccca
     5461 gtctttcact ttcgaagaag tagaatacct ggttaagggt ctgcgtaaca aattccaact
     5521 gaactgttac gtaaaaatca acaaaaacaa accgatcatc tacatcgatt ctatgtctta
     5581 cctgatcttc tacaacctga tcaaaccgta cctgatcccg cagatgatgt acaaactgcc
     5641 gaacactatc tcctccgaaa ctttcctgaa ataaccatgg gtatggacag ttttcccttt
     5701 gatatgtaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata
     5761 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt
     5821 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatact tactttgcat
     5881 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg
     5941 tagtgttttt cttagtccgt tatgtaggta ggaatctgat gtaatggttg ttggtatttt
     6001 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc
     6061 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt
     6121 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct
     6181 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat
     6241 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat
     6301 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt
     6361 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct
     6421 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct
     6481 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt
     6541 ttccctactg atgttcatca tctgaacgta ttggttataa gtgaacgata ccgtccgttc
     6601 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt
     6661 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac
     6721 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg
     6781 ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg
     6841 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc
     6901 tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata
     6961 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc
     7021 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa
     7081 ccctaaaggc ttaagtagca ccctcgcaag ctcggttgcg gccgcaatcg ggcaaatcgc
     7141 tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt tcgtgacatt
     7201 cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta caggcgcctt
     7261 ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac gggcttctca
     7321 gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt tcagcagttc
     7381 ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt cattcagact
     7441 ggctaatgca cccagtaagg cagcggtatc atcaacgggg tctgacgctc agtggaacga
     7501 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7561 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7621 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7681 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7741 ccccagtgct gcaa
//
