LOCUS       40924_1979        6755 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Integration vector mini-CTX2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6755)
  AUTHORS   Hoang TT, Kutchma AJ, Becher A, Schweizer HP.
  TITLE     A site-specific gene integration system for Pseudomonas aeruginosa
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6755)
  AUTHORS   Hoang TT, Kutchma AJ, Becher A, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-APR-1999) Microbiology, Colorado State University, 
            Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 6755)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6755)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-APR-1999) Microbiology, Colorado State University, Fort Collins,
            CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        827..845
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    858..965
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(974..992)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(1166..1213)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     oriT            complement(1420..1528)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    complement(2877..2893)
                     /label=lac operator
                     /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)."
     promoter        complement(2901..2930)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             complement(3174..4253)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4254..4331)
                     /label=lacI promoter
     misc_feature    4517..4657
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4843..5431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5511..6698)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(join(6746..6755,1..19))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
ORIGIN
        1 atcgatgata agctgtcaaa catgagaatt aattcatgtt ctttcctgcg ttatcccctg
       61 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      121 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
      181 ctctccccgc gcgttggccg attcattaat gcagcgaatt agcttcaaaa gcgctctgaa
      241 gttcctatac tttctagaga ataggaactt cggaatagga acttcaagat cccctgattc
      301 cctttgtcaa cagcaatgga ttttcaatgg gtcaacatga aatccggctg ttgccctggg
      361 acggcaggtt tgacggcagg cttccaggct ctctcgcaca cgctgggaac actctgggaa
      421 caaacgcgag tgaaaatgga cagtagggcc caggaataat gcggcctcca gcgaggttgg
      481 ggcgtttcat atgagttcga atctcaccgc ctccgccata taacctgctg ttttcattga
      541 tgtttccatt cgacccactc tcaggagtga acgatttggg aacaaactgg gaacgaggcg
      601 cagaaaaaag gggccccgca gggccccttt ttcgttgctt caggaaagtt tcagcgcctg
      661 attcagcaga cctacgatgt cggggccgtc cttgctgaat tagctttatg cttgtaaacc
      721 gttttgtgaa aaaattttta aaataaaaaa ggggacctct agggtcccca attaattagt
      781 aatataatct attaaaggtc attcaaaagg tcatccaccg gcgcgcaatt aaccctcact
      841 aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctctagaact agtggatccc
      901 ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag ggggggcccg
      961 gtacccaatt cgccctatag tgagtcgtat tacgcgcgcc ggtggatgac cttttgaatg
     1021 acctttaata gattatatta ctaattaatt ggggacccta gaggtcccct tttttatttt
     1081 aaaaattttt tcacaaaacg gtttacaagc ataaagctaa ttccattgag taagttttta
     1141 agcacatcag cttcaaaagc gctctgaagt tcctatactt tctagagaat aggaacttcg
     1201 gaataggaac ttcaagatcc ccaattcgct gcattaatga atcggccaac gcgcggggag
     1261 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1321 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1381 atcaggggat aacgcaggaa agaacatgct agagcttatc ggccagcctc gcagagcagg
     1441 attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc
     1501 gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc
     1561 tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa
     1621 aatcgctata atgaccccga agcagggtta tgcagcggaa agtatacctt aacctttttc
     1681 gttgcttcag gaaagtttca gcgcctgatt cagcagacct acgatgtcgg ggccgtcctt
     1741 gctgatccac ttggcgtaat gcttgaagat catcgccgtc gaggtgtgcc ccatctggtc
     1801 cgcgatccat tcgggagtgg cgatgccgct gctgagcatc tggctggcga aggtgtggcg
     1861 gcactgattg ggaccacgcg agcggacgcc cgcattcctg aggtggccat gccaccagcc
     1921 gttgcgcagc gtatccgacg tggagtacgc cgccccgcta gcactgttgt ggaacacgaa
     1981 gcgtacccgt tgaggctttc tggtccggtt gtcgcggtct atcacctcga tctgtacggg
     2041 cggaaggcgt ctggttagct tggcctgctg ctggagtgcg cgcagcgccg gggccagtag
     2101 ctggaccttc ctcgtcgatc ggcgagtctt ggtgaccttg tactggctgc gtacacgagc
     2161 gcggcggaaa accacggtcc ccgtgtccaa atcgacatct tcccaggcca gagctatggc
     2221 ctcgctgacg cgggggccgc tccagatcat gaactccgcc aggttcagct caccgatgcg
     2281 cgcggtttca gtaccgagga tgaggtcgat ttcctcacga gtgaatgggt ctgggtcatc
     2341 cgcatccgga agagtgatca cgatgccgtc tgttgggtcg tgcgcgctcc ggttccgagt
     2401 tcgatacagc cggaagatct gccgcaggtt gctgacgatc tctcgcaccg tcttgttgtg
     2461 cagcttgggc atgagcgtgt tctgcaccca gtcttgaata tccaggtggt cgatgctgtc
     2521 ggcctggctg tcaccccagc gaggtcggat gtgggtttcc acgcggctgg tgtagccgcg
     2581 gaagccgctg gcggcaatct ggttgcgttt gatgtcgagc cagaggtcga tgtaatggcc
     2641 caaggtgttg cttttcaccc gcggtgagtc cgggaagtgt cggctgtagc tgaagactcc
     2701 ttgcttgatc tcgtagttga tgatgccggc cagccgttcg gcattggcta tattctcggg
     2761 cgttgcgtcc cctggtatgg actccctaca cagctcgccc tggtagcgga agtagatcct
     2821 gatccgtttt ccgcgtacct cgactccgtc tgccatggtc tgtttcctgt gtgaaattgt
     2881 tatccgctca caattccaca cattatacga gccggatgat taattgtcaa cagctcattt
     2941 cagaatattt gccagaaccg ttatgatgtc ggcgcaaaaa acattatcca gaacgggagt
     3001 gcgccttgag cgacacgaat tatgcagtga tttacgacct gcacagccat accacagctt
     3061 ccgatggctg cctgacgcca gaagcattgg tgcaccgtgc agtcgatgat aagctgtcaa
     3121 acatgagaat taagatcaat tcgcgctaac tcacattaat tgcgttgcgc tcactgcccg
     3181 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3241 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     3301 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     3361 tgccccagca ggcgaaaatc ctgtttgatg gtggttgacg gcgggatata acatgagctg
     3421 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     3481 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     3541 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     3601 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     3661 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     3721 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     3781 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     3841 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     3901 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     3961 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     4021 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     4081 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     4141 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     4201 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     4261 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     4321 tcgatggtgt caacgtaaat gcatgccgct tcgccttcgc gcgcgaattg atctgctgcc
     4381 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4441 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     4501 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     4561 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     4621 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     4681 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4741 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5281 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5401 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5461 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatggtca ggtcgaggtg
     5521 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct
     5581 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc
     5641 agcggtccag tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc
     5701 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
     5761 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc
     5821 aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     5881 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     5941 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     6001 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     6061 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     6121 ggcatcggtc ggcgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     6181 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gtaaggagat ggcgcccaac
     6241 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     6301 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     6361 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agagaatcca caggacgggt
     6421 gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag caggactggg
     6481 cggcggccaa agcggtcgga cagtgctccg agaacgggtg cgcatagaaa ttgcatcaac
     6541 gcatatagcg ctagcagcac gccatagtga ctggcgatgc tgtcggaatg gacaatatcc
     6601 cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg
     6661 gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag
     6721 caatttaact gtgataaact accgcattaa agcta
//
