LOCUS       40924_27558        6861 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLAL1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6861)
  AUTHORS   Lacerda LA, Ferreira H.
  TITLE     Xanthomonas citri Expression Vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6861)
  AUTHORS   Lacerda LA, Ferreira H.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JAN-2015) Bioquimica e Microbiologia, UNESP, Av. 24A, 
            1515, Rio Claro, SP 13506900, Brazil
REFERENCE   3  (bases 1 to 6861)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6861)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-JAN-2015) Bioquimica e Microbiologia, UNESP, Av. 24A, 1515, Rio 
            Claro, SP 13506900, Brazil"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2452
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3239..3255
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3361..4236)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4263..4547
                     /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)"
     CDS             4582..4815
                     /codon_start=1
                     /label=ACP
                     /note="acyl carrier protein (ACP) from E. coli"
                     /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGADSLDTVELVM
                     ALEEEFDTEIPDEEAEKITTVQAAIDYINGHQA"
     CDS             4825..4902
                     /codon_start=1
                     /label=CBP
                     /note="calmodulin-binding peptide"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             4930..4950
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             4990..5163
                     /codon_start=1
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             5167..5337
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     primer_bind     complement(5378..5394)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5424..5442)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5463..5479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5487..5503)
                     /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(5511..5541)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5556..5577)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5851..5879
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6068..6598
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc
     1381 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg
     1441 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg
     1501 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag
     1561 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta
     1621 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg
     1681 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc
     1741 tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac
     1801 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
     1861 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
     1921 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
     1981 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     2041 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     2101 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     2161 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     2221 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     2281 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     2341 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     2401 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     2461 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     2521 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     2581 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga
     2641 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg
     2701 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     2761 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     2821 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag
     2881 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc
     2941 agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3001 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3061 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3121 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3181 gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc
     3241 tagaactagt ggatccccca aatctagagc ataatgtgcc tgtcaaatgg acgaagcagg
     3301 gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaatta
     3361 tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg
     3421 gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac
     3481 attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct
     3541 gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg
     3601 tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct
     3661 gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg
     3721 atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat
     3781 cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag
     3841 gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat
     3901 tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg
     3961 ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag
     4021 caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc
     4081 gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc
     4141 gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta
     4201 tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc cgccattcag
     4261 agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct
     4321 cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac
     4381 caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat
     4441 tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc
     4501 ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc cgtttttttg
     4561 ggctagcagg aggaattcac catgagcact atcgaagaac gcgttaagaa aattatcggc
     4621 gaacagctgg gcgttaagca ggaagaagtt accaacaatg cttctttcgt tgaagacctg
     4681 ggcgcggatt ctcttgacac cgttgagctg gtaatggctc tggaagaaga gtttgatact
     4741 gagattccgg acgaagaagc tgagaaaatc accaccgttc aggctgccat tgattacatc
     4801 aacggccacc aggcgtccat ggaaaagaga agatggaaaa agaatttcat agccgtctca
     4861 gcagccaacc gctttaagaa aatctcatcc tccggggcac ttgattatga tattccaact
     4921 actgctagcg agaatttgta ttttcagggt gagctcaaaa ccgcggctct tgcgcaacac
     4981 gatgaagccg tggacaacaa attcaacaaa gaacaacaaa acgcgttcta tgagatctta
     5041 catttaccta acttaaacga agaacaacga aacgccttca tccaaagttt aaaagatgac
     5101 ccaagccaaa gcgctaacct tttagcagaa gctaaaaagc taaatgatgc tcaggcgccg
     5161 aaagtagaca acaaattcaa caaagaacaa caaaacgcgt tctatgagat cttacattta
     5221 cctaacttaa acgaagaaca acgaaacgcc ttcatccaaa gtttaaaaga tgacccaagc
     5281 caaagcgcta accttttagc agaagctaaa aagctaaatg gtgctcaggc gccgaaagta
     5341 gacgcgaatt ccgcggggaa gtcaacctga agcttatcga taccgtcgac ctcgaggggg
     5401 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     5461 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5521 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5581 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5641 atcggccaac gcgcggggag aggcggtttg cgtattgggc gcatgcataa aaactgttgt
     5701 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     5761 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatgca tggacgcaca
     5821 ccgtggaaac ggatgaaggc acgaacccag ttgacataag cctgttcggt tcgtaaactg
     5881 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     5941 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt acagtctatg
     6001 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     6061 agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     6121 gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg accaagtcaa
     6181 atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag ccacctactc
     6241 ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga cattcatcgc
     6301 gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg ttctgcccag
     6361 gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg gcgagcaccg
     6421 gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca acgcgcttgg
     6481 tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg ctctctatac
     6541 aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta ccgccaccta
     6601 acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta aattgtcaca
     6661 acgccgccag gtggcacttt tcggggaaat gtgcgcgccc gcgttcctgc tggcgctggg
     6721 cctgtttctg gcgctggact tcccgctgtt ccgtcagcag cttttcgccc acggccttga
     6781 tgatcgcggc ggccttggcc tgcatatccc gattcaacgg ccccagggcg tccagaacgg
     6841 gcttcaggcg ctcccgaagg t
//
