LOCUS       40924_24523        6583 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHF5Ca, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6583)
  AUTHORS   Martins PMM., Oehlmeyer T, Amaral AM, Ferreira H.
  TITLE     A protein expression system for tandem affinity purification in 
            Xanthomonas axonopodis pv. citri
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6583)
  AUTHORS   Martins PMM., Oehlmeyer T, Amaral AM, Ferreira H.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-DEC-2008) Biologia, UNESP, Rua Cristovao Colombo, 
            2265, Sao Jose do Rio Preto, SP 15054000, Brazil
REFERENCE   3  (bases 1 to 6583)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6583)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-DEC-2008) Biologia, UNESP, Rua Cristovao Colombo, 2265, Sao Jose
            do Rio Preto, SP 15054000, Brazil"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(237..1289)
                     /codon_start=1
                     /gene="xylR"
                     /product="XylR"
                     /label=xylR
                     /note="xylose repressor form xylose operon"
                     /protein_id="ACL97676.1"
                     /translation="MTGLNKSTVSSQVNTLMKESMVFEIGQGQSSGGRRPVMLVFNKKA
                     GYSVGIDVGVDYINGILTDLEGTIVLDQYRHLESNSPEITKDILIDMIHHFITQMPQSP
                     YGFIGIGICVPGLIDKDQKIVFTPNSNWRDIDLKSSIQEKYNVSVFIENEANAGAYGEK
                     LFGAAKNHDNIIYVSISTGIGIGVIINNHLYRGVSGFSGEMGHMTIDFNGPKCSCGNRG
                     CWELYASEKALLKSLQTKEKKLSYQDIINLAHLNDIGTLNALQNFGFYLGIGLTNILNT
                     FNPQAVILRNSIIESHPMVLNSMRSEVSSRVYSQLGNSYELLPSSLGQNAPALGMSSIV
                     IDHFLDMITM"
     gene            complement(237..1289)
                     /gene="xylR"
                     /label=xylR
     CDS             1649..1726
                     /codon_start=1
                     /label=CBP
                     /note="calmodulin-binding peptide"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1754..1774
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             1814..1987
                     /codon_start=1
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1991..2161
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     CDS             2212..3032
                     /codon_start=1
                     /gene="Xac amy"
                     /product="truncated alpha amylase"
                     /label=Xac amy
                     /protein_id="ACL97680.1"
                     /translation="PGDVILHAFNWPYATVEARAKQIADAGYRKVLVAPAYRSEGSAWW
                     ARYQPQDIRLIDNPLGDTAAFKKMVQALASNGVETYADIVFNHMANEAATRSDLNYPGG
                     AVLSQYAANPGRYDSLRLFGTLQSNFLSASDFGPAQCISNYNDAYQVRNYRICGGGSDP
                     GLPDLLGNDWVVQQQRAYLQALKSMGVTGFRVDAAKHMTFDHLNRVFDAGIRSGVYVFG
                     EVITGGGTGNGDYDQFLAPYLQSTPHAAYDFPLFNAVRNAFAIGASMQQLP"
     gene            2212..3032
                     /gene="Xac amy"
                     /label=Xac amy
     primer_bind     complement(3042..3058)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3199..3627
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             3971..4762
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4783..5640
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5814..6402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gaattcttac
      241 attgtaatca tgtccagaaa atgatcaatc acaatggagg acattcctaa tgccggtgca
      301 ttctgtccta aggaagatgg caataattca tagctattgc ctaattggga ataaaccctt
      361 gatgatactt cacttctcat tgaatttaaa accataggat gcgattcaat tatgctattt
      421 cttaaaatta cggcttgtgg gttgaaagta tttagaatat tggtaaggcc tattcctaaa
      481 tagaatccaa aattttgtaa tgcatttaag gttccgatat cattcagatg ggcgaggttt
      541 atgatatctt gataggacag ttttttctct ttggtctgaa gagattttaa taaagccttc
      601 tctgaagcat acaattccca gcatcctcgg tttccgcaac tgcatttagg accattaaag
      661 tctattgtca tatgtcccat ttctccagag aagccgctta ctcctctata taaatgattg
      721 ttgataataa caccgatccc tattcctgtg ctgatactta cgtaaataat gttatcgtga
      781 ttttttgcag ctccaaatag tttttctcca tatgcgccag catttgcctc attttcaata
      841 aaaacagaca cattgtactt ctcttgtatc gaagatttta agtcaatatc tctccagttg
      901 gagttcggag tgaaaacaat tttttgatct ttatcaatga gtccaggcac gcaaatacct
      961 ataccaataa acccgtacgg agattggggc atttgcgtaa taaagtgatg aatcatatca
     1021 atcaaaatgt ctttcgttat ttctggagaa ttggattcca aatggcggta ttgatcaaga
     1081 acgattgttc cttcaaggtc tgttaaaatg ccattaatat aatccacacc aacatctatt
     1141 ccaacggagt atcctgcctt tttattaaaa acaagcatga caggtcttct tccgccactt
     1201 gattgtcctt gacctatttc aaataccata ctttctttca ttaacgtgtt tacctgtgat
     1261 gagacagttg atttatttaa tccagtcatt tcagataatt ttgctcttga aataggtgaa
     1321 tttttaagga tttcttttaa taataacttt tgatttactt ttttgacaaa ggtttgatca
     1381 gcgatatcca cttcatccac tccatttgtt taatctttaa attaagtatc aacatagtac
     1441 atagcgaatc ttccctttat taaatctaat gtgttcataa aaaactaaaa aaaatattga
     1501 aaatactgac gaggttatat aagatgaaaa taagttagtt tgtttaaaca acaaactaat
     1561 aggtgatgta cttactatat gaaataaaat gcatctgtat ttgaatgaat ttattaagct
     1621 tgggaaaagg aggtgaggat ccatggaaaa gagaagatgg aaaaagaatt tcatagccgt
     1681 ctcagcagcc aaccgcttta agaaaatctc atcctccggg gcacttgatt atgatattcc
     1741 aactactgct agcgagaatt tgtattttca gggtgagctc aaaaccgcgg ctcttgcgca
     1801 acacgatgaa gccgtggaca acaaattcaa caaagaacaa caaaacgcgt tctatgagat
     1861 cttacattta cctaacttaa acgaagaaca acgaaacgcc ttcatccaaa gtttaaaaga
     1921 tgacccaagc caaagcgcta accttttagc agaagctaaa aagctaaatg atgctcaggc
     1981 gccgaaagta gacaacaaat tcaacaaaga acaacaaaac gcgttctatg agatcttaca
     2041 tttacctaac ttaaacgaag aacaacgaaa cgccttcatc caaagtttaa aagatgaccc
     2101 aagccaaagc gctaaccttt tagcagaagc taaaaagcta aatggtgctc aggcgccgaa
     2161 agtagacgcg aattccgcgg ggaagtcaac ctgaagcttg atctagaatg acccggggat
     2221 gtcatcctgc acgccttcaa ctggccgtat gccaccgtgg aagcgcgcgc caagcagatt
     2281 gccgatgccg gctatcgcaa ggtattggtc gcccccgcgt accgctccga aggaagtgcc
     2341 tggtgggcgc gctatcaacc ccaggacatc cgcctgatcg acaatccgct cggcgacacc
     2401 gctgcattca agaagatggt ccaggcgctg gccagcaacg gcgtggaaac ctatgccgac
     2461 atcgtgttca accacatggc caacgaagcg gccacacgct cggacctcaa ttacccaggc
     2521 ggcgcggtcc tgtcgcagta cgcggcaaac cccggccgtt acgattcgct gcgcttgttc
     2581 ggcacgctgc agtcaaactt cctcagcgcc agcgatttcg gcccggcgca gtgcatcagc
     2641 aactacaacg acgcttacca ggtgcgcaat taccgtatct gcggcggcgg cagcgatcca
     2701 ggcctgccgg atttgctggg caacgactgg gtggtacagc agcagcgcgc ctatctgcag
     2761 gcgctcaaga gcatgggcgt gaccggcttc cgggtggatg cggccaagca catgacgttc
     2821 gaccatctca atcgcgtctt cgatgccggc atccgatcgg gtgtgtatgt cttcggcgaa
     2881 gtgatcaccg gcggcggtac gggcaatggc gattacgacc agttcctggc gccgtacctg
     2941 caatccacgc cgcatgcggc ctacgacttt ccgctgttca atgcggtgcg caacgcgttc
     3001 gccattggcg ccagcatgca gcagctcccg ggaagcttgg cactggccgt cgttttacaa
     3061 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     3121 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     3181 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3241 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3301 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3361 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3421 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3481 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3541 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3601 tgatttaaca aaaatttaac gcgaatttta acaaaattca gggcgcaagg gctgctaaag
     3661 gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag
     3721 ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag
     3781 tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt
     3841 gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt
     3901 cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgagg
     3961 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4021 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4081 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4141 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4201 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4261 gccggggcag gatctcctgt catcccacct tgctcctgcc gagaaagtat ccatcatggc
     4321 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4381 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4441 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4501 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4561 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4621 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4681 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4741 tcgccttctt gacgagttct tctgaattga aaaaggaaga gtatgagtat tcaacatttc
     4801 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4861 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4921 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4981 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5041 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5101 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5161 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5221 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5281 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5341 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5401 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5461 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5521 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5581 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5641 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5701 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5761 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5821 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5881 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5941 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6001 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6061 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6121 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6181 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6241 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6301 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6361 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6421 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6481 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6541 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
