LOCUS       Exported                8572 bp ds-DNA     circular SYN 06-OCT-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pS238D1::sptse5-CT_phoA-lacZ-alpha
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8572)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 6, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8572
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(19..984)
                     /codon_start=1
                     /gene="xylS"
                     /product="XylS regulator encoded by the Pseudomonas putida
                     TOL plasmid pWWO"
                     /label=XylS
                     /note="stimulates transcription from the Pm promoter in the
                     presence of aromatic compounds such as m-toluate or 
                     benzoate"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     promoter        1884..1964
                     /label=Pm promoter
                     /note="The bacterial Pm promoter is activated by XylS in
                     the presence of benzoate or m-toluate (Marques et al., 
                     1999)."
     CDS             2017..3099
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     sig_peptide     3134..3199
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /note="/codon_start=1"
                     /note="/translation=MKYLLPTAAAGLLLLAAQPAMA"
     CDS             3209..3655
                     /codon_start=1
                     /label=Tse5-CT
                     /translation="IIPLVVIGAFAARAAIGAALGAGIELGMQTGKQVLGQMKDNWDSD
                     RDLTDIKWKCIDINWKHVGASAAIGTVAPGMLSTGKTVVQSAKAIRTLSGQAANTANRA
                     AKLAARKAAHADTIKKAVATQAAWQTGKQIVKCPLKDEEEECPPQ"
     CDS             3671..5188
                     /codon_start=1
                     /label=PhoA_LacZalpha
                     /translation="TPEMPVLENRAAQGDITAPGGARRLTGDQTAALRDSLSDKPAKNI
                     ILLIGDGMGDSEITAARNYAEGAGGFFKGIDALPLTGQYTHYALNKKTGKPDYVTDSAA
                     SATAWSTGVKTYNGALGVDIHEKDHPTILEMAKAAGLATGNVSTAELQDATPAALVAHV
                     TSRKCYGPSATSEKCPGNALEKGGKGSITEQLLNARADVTLGGGAKTFAETATAGEWQG
                     KTLREQAQARGYQLVSDAASLNSVTEANQQKPLLGLFADGNMPVRWLGPKATYHGNIDK
                     PAVTCTPNPQRNDSVPTLAQMTDKAIELLSKNEKGFFLQVEGASIDKQDHAANPCGQIG
                     ETVDLDEAVQRALEFAKKEGNTLVIVTADHAHASQIVAPDTKAPGLTQALNTKDGAVMV
                     MSYGNSEEDSQEHTGSQLRIAAYGPHAANVVGLTDQTDLFYTMKAALGLKTDSLAVVLQ
                     RRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     primer_bind     complement(5026..5042)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(5305..5332)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(5348..5418)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5604..5698
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             5830..6645
                     /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"
     oriT            6812..6920
                     /label=incP origin of transfer
                     /note="incP origin of transfer"
     CDS             complement(6931..7593)
                     /codon_start=1
                     /product="replication protein for the broad-host-range
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      7594..8365
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(8480..8566)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaac gttcgtaatc aagccacttc ctttttgcat tgacgcaggg tgtcggaagg
       61 caactcgccg aacgcgctcc tatagttttc agcgaagcgt cccaaatgta agaagccgta
      121 gtctagggct atctcagtta tactacgcac attggcactg ggatcgttca agcaggcgcg
      181 gatgctttcg agcttgcggt tgcggatgta gttcttcggc gtggtgccgg cgtgcttctc
      241 gaacaaattg tagagcgagc gtggactcat catcgccagc tccgctaacc gctcaaggct
      301 gatattccgt ttgagattct cctcaatgaa ttgaacgact cgctcgaaag acgggttacc
      361 tttgctgaaa atttcacggc tgacattgct gcccagcatt tcgagcagct tggaagcgat
      421 gatccccgca tagtgctctt ggacccgagg catcgacttt gtatgttccg cttcgtcaca
      481 aactaacccg agtagattga taaagccatc gagttgctgg agattgtgtc gcgcggcgaa
      541 acggataccc tccctcggct tgtgccaatt gttgtcactg cacgcccgat caaggaccac
      601 tgagggcaat ttaacgataa atttctcgca atcttctgaa taggtcaggt cggcttggtc
      661 atccggattg agcagcaata gttcgcccgg cgcaaaatag tgctcctggc catggccacg
      721 ccacaggcaa tggcctttga gtattatttg cagatgataa caggtctcta atccaggcga
      781 gattaccctc acgctaccgc cgtagctgat tcgacacaga tcgaggcatc cgaagattct
      841 gtggtgcagc ctgcctgccg ggcgcccgcc cttgggcagg cgaatagagt gcgtaccgac
      901 atactggtta acataatcgg agactgcata gggctcggcg tggacgaaga tctgactttt
      961 ctcgttcaat aagcaaaaat ccatagttca cggttctctt attttaatgt gggctgcttg
     1021 gtgtgatgta gaaaggcgcc aagtcgatga aaatgcatct cgacgtgatg cgtatacggg
     1081 ttacccccat tgccacgttg cgccatcctt tttgcaatca gtgaccactt ttccaagcaa
     1141 aaataacgcc aagcagaacg aagacgttct ttttaagaag cgagaacacc agaagttcgt
     1201 gctgtcgggg catggggcga cgaattggcg gataaagggg atctgctgga tattacggcc
     1261 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1321 gcccgcctga tgaatgctca tccgtaatta cgtatggcaa tgaaagacgg tgagctggtg
     1381 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1441 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1501 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1561 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1621 gacaacttct tcgcccccgt tttcaccatg catgggaatt agcttgatct gaccaacgac
     1681 cggtagcgga gctatccaac ggcggtatac caggaaaaca cacagcaggt acatcagaac
     1741 agtaccatga ctgaagaaca aatagttttt tcctgatcca taaagcagaa cggcctgctc
     1801 catgacaaat ctggctcccc aactaatgcc ccatgcagcc agcataacca gcataaagtg
     1861 cagtgtccgg tttgataggg ataagtccag ccttgcaaga agcggataca ggagtgcaaa
     1921 aaatggctat ctctagtaag gcctacccct taggctttat gcaacagaaa caataataat
     1981 ggagtcatga ccatgcctag gaggaaaaaa catatggtga aaccagtaac gttatacgat
     2041 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     2101 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     2161 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     2221 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     2281 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     2341 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     2401 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     2461 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
     2521 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     2581 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     2641 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     2701 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     2761 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     2821 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc
     2881 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     2941 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     3001 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3061 ctggcacgac aggtttcccg actggaaagc gggcagtaac atcaccacca tcattaggat
     3121 ggtggtgatg ataatgaagt acctgctgcc gaccgcggcg gcgggtctgc tgctgctggc
     3181 ggcgcagccg gcgatggcga tgggcctgat cattccgctg gttgtgatcg gtgcgtttgc
     3241 ggcgcgtgcg gcgattggtg cggcgctggg tgcgggcatt gagctgggta tgcagaccgg
     3301 caaacaagtg ctgggtcaga tgaaggataa ctgggacagc gatcgtgacc tgaccgatat
     3361 caagtggaaa tgcatcgaca ttaactggaa acatgttggt gcgagcgcgg cgattggtac
     3421 cgttgcgccg ggcatgctga gcaccggtaa aaccgtggtt cagagcgcga aggcgattcg
     3481 taccctgagc ggtcaggcgg cgaacaccgc gaaccgtgcg gcgaaactgg cggcgcgtaa
     3541 ggcggcgcac gcggatacca tcaagaaagc ggtggcgacc caagcggcgt ggcagaccgg
     3601 caagcaaatt gttaaatgcc cgctgaagga cgaggaagag gaatgcccgc cgcaagggag
     3661 ctcggcccgg acaccagaaa tgcctgttct ggaaaaccgg gctgctcagg gcgatattac
     3721 tgcacccggc ggtgctcgcc gtttaacggg tgatcagact gccgctctgc gtgattctct
     3781 tagcgataaa cctgcaaaaa atattatttt gctgattggc gatgggatgg gggactcgga
     3841 aattactgcc gcacgtaatt atgccgaagg tgcgggcggc ttttttaaag gtatagatgc
     3901 cttaccgctt accgggcaat acactcacta tgcgctgaat aaaaaaaccg gcaaaccgga
     3961 ctacgtcacc gactcggctg catcagcaac cgcctggtca accggtgtca aaacctataa
     4021 cggcgcgctg ggcgtcgata ttcacgaaaa agatcaccca acgattctgg aaatggcaaa
     4081 agccgcaggt ctggcgaccg gtaacgtttc taccgcagag ttgcaggatg ccacgcccgc
     4141 tgcgctggtg gcacatgtga cctcgcgcaa atgctacggt ccgagcgcga ccagtgaaaa
     4201 atgtccgggt aacgctctgg aaaaaggcgg aaaaggatcg attaccgaac agctgcttaa
     4261 cgctcgtgcc gacgttacgc ttggcggcgg cgcaaaaacc tttgctgaaa cggcaaccgc
     4321 tggtgaatgg cagggaaaaa cgctgcgtga acaggcacag gcgcgtggtt atcagttggt
     4381 gagcgatgct gcctcactga attcggtgac ggaagcgaat cagcaaaaac ccctgcttgg
     4441 cctgtttgct gacggcaata tgccagtgcg ctggctagga ccgaaagcaa cgtaccatgg
     4501 caatatcgat aagcccgcag tcacctgtac gccaaatccg caacgtaatg acagtgtacc
     4561 aaccctggcg cagatgaccg acaaagccat tgaattgttg agtaaaaatg agaaaggctt
     4621 tttcctgcaa gttgaaggtg cgtcaatcga taaacaggat catgctgcga atccttgtgg
     4681 gcaaattggc gagacggtcg atctcgatga agccgtacaa cgggcgctgg aattcgctaa
     4741 aaaggagggt aacacgctgg tcatagtcac cgctgatcac gcccacgcca gccagattgt
     4801 tgcgccggat accaaagctc cgggcctcac ccaggcgcta aataccaaag atggcgcagt
     4861 gatggtgatg agttacggga actccgaaga ggattcacaa gaacataccg gcagtcagtt
     4921 gcgtattgcg gcgtatggcc cgcatgccgc caatgttgtt ggactgaccg accagaccga
     4981 tctcttctac accatgaaag ccgctctggg gctgaaaacg gattcactgg ccgtcgtttt
     5041 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5101 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5161 gcgcagcctg aatggcgaat ggcgctaatg aggtacccgg ggatcctcta gagtcgacct
     5221 gcaggcatgc aagcttgcgg ccgcgtcgtg actgggaaaa ccctggcgca aagcccgccg
     5281 aaaggcgggc ttttctgtta taaacgcaga aaggcccacc cgaaggtgag ccagtgtgac
     5341 tctagtagag agcgttcacc gacaaaaaca gataaaacga aaggcccagt ctttcgactg
     5401 agcctttcgt tttatttgat gcctggctcg agctcgagaa aaaaagcccg gacgactgtt
     5461 cgggcttgtc tttttatatg ttggaaaatc agtggcaatg caatggccca acagaaaatg
     5521 gtgaaaccag taacgttata ctgaatctaa gtatcaatgt gagcgctcac aagtcaacac
     5581 tctttttgat aaaactagtc ttggactcct gttgatagat ccagtaatga cctcagaact
     5641 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc
     5701 aggggtcccc aataattacg atttaaattt gtgtctcaaa atctctgatg ttacattgca
     5761 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     5821 aggggtgtta tgagccatat tcagcgtgaa acgagctgta gccgtccgcg tctgaacagc
     5881 aacatggatg cggatctgta tggctataaa tgggcgcgtg ataacgtggg tcagagcggc
     5941 gcgaccattt atcgtctgta tggcaaaccg gatgcgccgg aactgtttct gaaacatggc
     6001 aaaggcagcg tggcgaacga tgtgaccgat gaaatggtgc gtctgaactg gctgaccgaa
     6061 tttatgccgc tgccgaccat taaacatttt attcgcaccc cggatgatgc gtggctgctg
     6121 accaccgcga ttccgggcaa aaccgcgttt caggtgctgg aagaatatcc ggatagcggc
     6181 gaaaacattg tggatgcgct ggccgtgttt ctgcgtcgtc tgcatagcat tccggtgtgc
     6241 aactgcccgt ttaacagcga tcgtgtgttt cgtctggccc aggcgcagag ccgtatgaac
     6301 aacggcctgg tggatgcgag cgattttgat gatgaacgta acggctggcc ggtggaacag
     6361 gtgtggaaag aaatgcataa actgctgccg tttagcccgg atagcgtggt gacccacggc
     6421 gattttagcc tggataacct gattttcgat gaaggcaaac tgattggctg cattgatgtg
     6481 ggccgtgtgg gcattgcgga tcgttatcag gatctggcca ttctgtggaa ctgcctgggc
     6541 gaatttagcc cgagcctgca aaaacgtctg tttcagaaat atggcattga taatccggat
     6601 atgaacaaac tgcaatttca tctgatgctg gatgaatttt tctaataatt aattggaccg
     6661 cggtccgcgc gttgtccttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
     6721 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
     6781 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
     6841 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
     6901 ctgctctgcg aggctggccg taggccggcc ctaccggcgc ggcagcgtta cccgtgtcgg
     6961 cggctccaac ggctcgccat cgtccagaaa acacggctca tcgggcatcg gcaggcgctg
     7021 ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg gttccatgcc
     7081 cggaatgccg ggctggctgg gcggctcctc gccggggccg gtcggtagtt gctgctcgcc
     7141 cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt cgtcctggtc
     7201 gtcgtgatca accaccacgg cggcactgaa caccgacagg cgcaactggt cgcggggctg
     7261 gccccacgcc acgcggtcat tgaccacgta ggccgacacg gtgccggggc cgttgagctt
     7321 cacgacggag atccagcgct cggccaccaa gtccttgact gcgtattgga ccgtccgcaa
     7381 agaacgtccg atgagcttgg aaagtgtctt ctggctgacc accacggcgt tctggtggcc
     7441 catctgcgcc acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg caataagccc
     7501 ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct tgttcttggc
     7561 ttgaatgccg atttctctgg actgcgtggc catgcttatc tccatgcggt aggggtgccg
     7621 cacggttgcg gcaccatgcg caatcagctg caacttttcg gcagcgcgac aacaattatg
     7681 cgttgcgtaa aagtggcagt caattacaga ttttctttaa cctacgcaat gagctattgc
     7741 ggggggtgcc gcaatgagct gttgcgtacc cccctttttt aagttgttga tttttaagtc
     7801 tttcgcattt cgccctatat ctagttcttt ggtgcccaaa gaagggcacc cctgcggggt
     7861 tcccccacgc cttcggcgcg gctccccctc cggcaaaaag tggcccctcc ggggcttgtt
     7921 gatcgactgc gcggccttcg gccttgccca aggtggcgct gcccccttgg aacccccgca
     7981 ctcgccgccg tgaggctcgg ggggcaggcg ggcgggcttc gcccttcgac tgcccccact
     8041 cgcataggct tgggtcgttc caggcgcgtc aaggccaagc cgctgcgcgg tcgctgcgcg
     8101 agccttgacc cgccttccac ttggtgtcca accggcaagc gaagcgcgca ggccgcaggc
     8161 cggaggcttt tccccagaga aaattaaaaa aattgatggg gcaaggccgc aggccgcgca
     8221 gttggagccg gtgggtatgt ggtcgaaggc tgggtagccg gtgggcaatc cctgtggtca
     8281 agctcgtggg caggcgcagc ctgtccatca gcttgtccag cagggttgtc cacgggccga
     8341 gcgaagcgag ccagccggtg gccgctcgcg gccatcgtcc acatatccac gggctggcaa
     8401 gggagcgcag cgaccgcgca gggcgaagcc cggagagcaa gcccgtaggg ggggcgcgcc
     8461 cagctgtcta gggcggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     8521 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ct
//
