LOCUS       40924_48568        6000 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pZS4Int-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6000)
  AUTHORS   Lutz R, Bujard H.
  TITLE     Independent and tight regulation of transcriptional units in 
            Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 
            regulatory elements
  JOURNAL   Nucleic Acids Res. 25 (6), 1203-1210 (1997)
  PUBMED    9092630
REFERENCE   2  (bases 1 to 6000)
  AUTHORS   Lutz R, Bujard H.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-AUG-1996) Zentrum fuer Molekulare Biologie, University
            of Heidelberg, Im Neuenheimer Feld 282, Heidelberg 69120, Germany
REFERENCE   3  (bases 1 to 6000)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6000)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "1997"; volume: "25"; issue: "6"; pages: 
            "1203-1210"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-AUG-1996) Zentrum fuer Molekulare Biologie, University of 
            Heidelberg, Im Neuenheimer Feld 282, Heidelberg 69120, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        120..197
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             198..1277
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             1454..2074
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline repressor TetR"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNKAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     protein_bind    complement(2174..2405)
                     /label=lambda attP
                     /note="integrase from phage lambda"
     terminator      2440..2526
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3064..4011)
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(4059..4281)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4773..4867)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4914..5702)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATYDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
       61 ccctttcgtc ttcacctcga caattcgcgc gcgaaggcga agcggcatgc atttacgttg
      121 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
      181 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
      241 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
      301 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
      361 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
      421 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
      481 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
      541 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
      601 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
      661 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
      721 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
      781 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
      841 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
      901 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
      961 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     1021 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     1081 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     1141 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     1201 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     1261 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagcgcga attgtcgagg
     1321 gaaatcataa aaaatttatt tgctttcagg aaaatttttc tgtataatag attcataaat
     1381 ttgagagaag agtttaaata tggctggttc tcgcagaaag aaacatatcc atgaaatccc
     1441 gccccgaatt catatgtcta gattagataa aagtaaagtg attaacagcg cattagagct
     1501 gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt
     1561 agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc
     1621 cattgagatg ttagataggc accatactca cttttgccct ttagaagggg aaagctggca
     1681 agatttttta cgtaataagg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg
     1741 agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca
     1801 attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc
     1861 tgtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga
     1921 agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt
     1981 atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg
     2041 attagaaaaa caacttaaat gtgaaagtgg gtcttaagct ttctagatag ctgagtagcc
     2101 tagagtgcgc gacaggtttg atgacaaaaa attagcgcaa gaagacaaaa atcaccttgc
     2161 gctaatgctc tgttacaggt cactaatacc atctaagtag ttgattcata gtgactgcat
     2221 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     2281 gatatttata tcattttacg tttctcgttc agctttttta tactaagttg gcattataaa
     2341 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2401 tatttgattt caattttgtc ccactccccc tagaggcatc aaataaaacg aaaggctcag
     2461 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     2521 acaaatccgc cgccctagac ctagggtacg ggttttgctg cccgcaaacg ggctgttctg
     2581 gtgttgctag tttgttatca gaatcgcaga tccggcttca ggtttgccgg ctgaaagcgc
     2641 tatttcttcc agaattgcca tgattttttc cccacgggag gcgtcactgg ctcccgtgtt
     2701 gtcggcagct ttgattcgat aagcagcatc gcctgtttca ggctgtctat gtgtgactgt
     2761 tgagctgtaa caagttgtct caggtgttca atttcatgtt ctagttgctt tgttttactg
     2821 gtttcacctg ttctattagg tgttacatgc tgttcatctg ttacattgtc gatctgttca
     2881 tggtgaacag ctttaaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt
     2941 acaccgtttt catctgtgca tatggacagt tttccctttg atatctaacg gtgaacagtt
     3001 gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc
     3061 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
     3121 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
     3181 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
     3241 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
     3301 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
     3361 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
     3421 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
     3481 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
     3541 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
     3601 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
     3661 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
     3721 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
     3781 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
     3841 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
     3901 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
     3961 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
     4021 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     4081 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     4141 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     4201 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     4261 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     4321 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     4381 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     4441 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     4501 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     4561 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     4621 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     4681 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     4741 ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg
     4801 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     4861 aggagtccaa gcgagctcga tatccgtcgg cttgaacgaa ttgttagaca ttatttgccg
     4921 actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc
     4981 gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc
     5041 tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt
     5101 ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg
     5161 ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga
     5221 tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta
     5281 tgttctcttg cttttgtcag caagatagcc agatcaatgt cgtacgtggc tggctcgaag
     5341 atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga
     5401 taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc
     5461 tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt
     5521 gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg
     5581 ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc
     5641 tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc
     5701 atgatgttta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctccat
     5761 aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac
     5821 tgtaccccaa aaaaacatgt cataacaaga agccatgaaa accgccactg cgccgttacc
     5881 accgctgcgt tcggtcaagg ttgtggacca gttgcgtgac ggcagttacg ctacttgcat
     5941 tacagcttac gaaccgaacg aggcttatgt ccactgggtt cgtgccttca tccggatatc
//
