LOCUS       40924_24528        6651 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHF5Na, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6651)
  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  JOURNAL   Unpublished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 6651)
  AUTHORS   Martins PMM., Oehlmeyer T, Amaral AM, Ferreira H.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-DEC-2008) Biologia, UNESP, Rua Cristovao Colombo, 
            2265, 15054000, SP, Brazil
REFERENCE   3  (bases 1 to 6651)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6651)
  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            "Unpublished"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, 
            15054000, SP, Brazil"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6651
                     /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 from xylose operon"
                     /protein_id="ACM16581.1"
                     /translation="MTGLNKSTVSSQVNTLMKESMVFEIGQGQSSGGRRPVMLVFNKKA
                     GYSVGIDVGVDYINGILTDLEGTIVLDQYRHLESNSPEITKDILIDMIHHFITQMPQSP
                     YGFIGIGICVPGLIDKDQKIVFTPNSNWRDIDLKSSIQEKYNVSVFIENEANAGAYGEK
                     LFGAAKNHDNIIYVSISTGIGIGVIINNHLYRGVSGFSGEMGHMTIDFNGPKCSCGNRG
                     CWELYASEKALLKSLQTKEKKLSYQDIINLAHLNDIGTLNALQNFGFYLGIGLTNILNT
                     FNPQAVILRNSIIESHPMVLNSMRSEVSSRVYSQLGNSYELLPSSLGQNAPALGMSSIV
                     IDHFLDMITM"
     gene            complement(237..1289)
                     /gene="xylR"
                     /label=xylR
     protein_bind    complement(1646..1679)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1712..1885
                     /codon_start=1
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1889..2059
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             2096..2116
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             2123..2200
                     /codon_start=1
                     /label=CBP
                     /note="calmodulin-binding peptide"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2201..2215
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     primer_bind     complement(2221..2237)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2280..3100
                     /codon_start=1
                     /gene="Xac amy"
                     /product="truncated alpha-amylase"
                     /label=Xac amy
                     /protein_id="ACM16584.1"
                     /translation="PGDVILHAFNWPYATVEARAKQIADAGYRKVLVAPAYRSEGSAWW
                     ARYQPQDIRLIDNPLGDTAAFKKMVQALASNGVETYADIVFNHMANEAATRSDLNYPGG
                     AVLSQYAANPGRYDSLRLFGTLQSNFLSASDFGPAQCISNYNDAYQVRNYRICGGGSDP
                     GLPDLLGNDWVVQQQRAYLQALKSMGVTGFRVDAAKHMTFDHLNRVFDAGIRSGVYVFG
                     EVITGGGTGNGDYDQFLAPYLQSTPHAAYDFPLFNAVRNAFAIGASMQQLP"
     gene            2280..3100
                     /gene="Xac amy"
                     /label=Xac amy
     primer_bind     complement(3110..3126)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3267..3695
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             4039..4830
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4851..5708
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5882..6470
                     /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 ccatgataac ttcgtatagc atacattata cgaagttatc
     1681 catggcaggc cttgcgcaac acgatgaagc cgtggacaac aaattcaaca aagaacaaca
     1741 aaacgcgttc tatgagatct tacatttacc taacttaaac gaagaacaac gaaacgcctt
     1801 catccaaagt ttaaaagatg acccaagcca aagcgctaac cttttagcag aagctaaaaa
     1861 gctaaatgat gctcaggcgc cgaaagtaga caacaaattc aacaaagaac aacaaaacgc
     1921 gttctatgag atcttacatt tacctaactt aaacgaagaa caacgaaacg ccttcatcca
     1981 aagtttaaaa gatgacccaa gccaaagcgc taacctttta gcagaagcta aaaagctaaa
     2041 tgatgctcag gcgccgaaag tagacgcgaa ttgtgatata cctacaactg cttctgaaaa
     2101 tttatatttt caaggtgaac taaagagaag atggaaaaag aatttcatag ccgtctcagc
     2161 agccaaccgc tttaagaaaa tctcatcctc cggggcactt gatgatgacg ataagcttat
     2221 cgataccgtc gacctcgagg gggggcccgg tacccaattc gccctatagt ctagaatgac
     2281 ccggggatgt catcctgcac gccttcaact ggccgtatgc caccgtggaa gcgcgcgcca
     2341 agcagattgc cgatgccggc tatcgcaagg tattggtcgc ccccgcgtac cgctccgaag
     2401 gaagtgcctg gtgggcgcgc tatcaacccc aggacatccg cctgatcgac aatccgctcg
     2461 gcgacaccgc tgcattcaag aagatggtcc aggcgctggc cagcaacggc gtggaaacct
     2521 atgccgacat cgtgttcaac cacatggcca acgaagcggc cacacgctcg gacctcaatt
     2581 acccaggcgg cgcggtcctg tcgcagtacg cggcaaaccc cggccgttac gattcgctgc
     2641 gcttgttcgg cacgctgcag tcaaacttcc tcagcgccag cgatttcggc ccggcgcagt
     2701 gcatcagcaa ctacaacgac gcttaccagg tgcgcaatta ccgtatctgc ggcggcggca
     2761 gcgatccagg cctgccggat ttgctgggca acgactgggt ggtacagcag cagcgcgcct
     2821 atctgcaggc gctcaagagc atgggcgtga ccggcttccg ggtggatgcg gccaagcaca
     2881 tgacgttcga ccatctcaat cgcgtcttcg atgccggcat ccgatcgggt gtgtatgtct
     2941 tcggcgaagt gatcaccggc ggcggtacgg gcaatggcga ttacgaccag ttcctggcgc
     3001 cgtacctgca atccacgccg catgcggcct acgactttcc gctgttcaat gcggtgcgca
     3061 acgcgttcgc cattggcgcc agcatgcagc agctcccggg aagcttggca ctggccgtcg
     3121 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3181 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3241 agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg
     3301 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3361 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3421 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3481 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3541 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3601 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3661 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaattcagg gcgcaagggc
     3721 tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg
     3781 aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta
     3841 gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa
     3901 ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg
     3961 atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca
     4021 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4081 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4141 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4201 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4261 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4321 ggcgaagtgc cggggcagga tctcctgtca tcccaccttg ctcctgccga gaaagtatcc
     4381 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4441 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4501 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4561 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4621 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4681 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4741 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4801 gccttctatc gccttcttga cgagttcttc tgaattgaaa aaggaagagt atgagtattc
     4861 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4921 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4981 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5041 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5101 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5161 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5221 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5281 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5341 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5401 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5461 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5521 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5581 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5641 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5701 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5761 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5821 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5881 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5941 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6001 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6061 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6121 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6181 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6241 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6301 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6361 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6421 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6481 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6541 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6601 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
