LOCUS       sequence-254660-        7746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Acinetobacter baumannii PirA in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7746)
  AUTHORS   Moynie L, Luscher A, Rolo D, Pletzer D, Tortajada A, Weingart H, 
            Braun Y, Page MG, Naismith JH, Kohler T
  TITLE     Structure and Function of the PiuA and PirA Siderophore-Drug 
            Receptors from Pseudomonas aeruginosa and Acinetobacter baumannii.
  JOURNAL   Antimicrob Agents Chemother. 2017 Mar 24;61(4). pii: AAC.02531-16. 
            doi: 10.1128/AAC.02531-16. Print 2017 Apr.
  PUBMED    28137795
REFERENCE   2  (bases 1 to 7746)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Antimicrob 
            Agents Chemother."; date: "2017-03-24"; volume: "61(4). pii: 
            AAC.02531-16. doi"; pages: " 10.1128/AAC.02531-16. Print 2017 Apr"
COMMENT     Sequence Label: pTAMAHISTEV_Ab-PirA
FEATURES             Location/Qualifiers
     source          1..7746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     RBS             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             151..171
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             193..213
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2459..2476
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2529..2547)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2543..2590
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2627..3082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2714..2733)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2924..2945
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3108..3212
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3213..3281,3282..4073)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3431..3450)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4244..4832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4733..4752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4986..5003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5018..5157
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5083..5102)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5243..5265
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5259..5450)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6259..7341)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     7303..7322
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7342..7419)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7625..7644
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(7728..7746,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7728..7746
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatatacata tgcgctatat ccgacagtta tgctgtgtat ccttactctg cttaagcgga
      121 tctgccgccg ccgcgaacgt ccgtctacag caccatcacc atcaccatca cgattacgat
      181 atcccaacga ccgaaaacct gtattttcag ggcgccatgg cacaaaatga gcaagaacaa
      241 gctgaacaaa cattagaaaa gcctgctgaa cctgtgaaat tggaaacaat ttttgtaaca
      301 gcagaagagc aagtgaagca atcgctcggt gtatcggtta ttaccaaaga agatttagaa
      361 aaactaccag ttcgtaatga tatttctgac tatgtacgtc gtatgccagg tgtcaacctg
      421 acaggcaata gtgctactgg gcagcgtggt aataatagac aaattgatat tcgcggaatg
      481 ggccctgaaa acacgcttat tttagtggat ggtaagccaa ttaactctcg taattcagtc
      541 cgttatggct ggaaaggaga gcgtgataca cgaggcgact caaactgggt accagcagaa
      601 gccatcgagt cgattgaagt tttacgcgga ccagcagctg ctcgatatgg ttctggtgct
      661 gcgggtggtg tagttaacat cattactaaa aaagtgacta atgaaactca tggttcagta
      721 gaattttata cttcacagcc tgaagactcc aaagaaggtt catcaaatcg tgttggtttt
      781 aacgtaagtg ggccactaat taaggacgtt ttatcttatc gtttatatgg taattataat
      841 aaaacagaag ctgacgatgt tgatattaat aaatcaattg gtagtaccgc agctggccgt
      901 gaaggtgtta aaaataaaga tatttcaggc cgtttagctt ggcaggcaac agaccagcaa
      961 actgtcttgc tcgatatttc ttctagcaaa caaggtaata tttattctgg tgactctcag
     1021 ttaaatgcaa atgctgaagc ggatgcgatt ctttcacagc tgattggtaa agaaaccaat
     1081 accatgtatc gtgatagcta tgcattaacg cacgaaggtg attggtcttg gggtaagagt
     1141 aagttagttg ctcaatatga taagacccat aacaaacgtc tacctgaagg cttggcggga
     1201 agtgtagaag gaaaaattaa taatcttgat gataaagcca cttcgcgttt agaaactctt
     1261 cgctttaacg gcgaggctaa tattcctttt gaatactatt taccccaagt attaactgta
     1321 ggtaccgaat gggttgaaga cagatttaaa gataatgtct cgacaactca aggtaaagac
     1381 agcagtggtt caggttatgg cgatcaatta gcgaaaggtg atcgtagtaa aatggagtca
     1441 cgtattgctt ctgcatatat tgaagataac ctgaaagtta cagacagcac agatgttgta
     1501 ttaggtttac gttttgatga ccatagtaaa tctggttcta attggagccc aagcttaaat
     1561 attactcaaa aactcaatga ttatttcact ttaaaaggtg gggtagcaaa agcttataaa
     1621 gcaccaaata tgtatcaaaa tgccgaaggg tatttattaa gtacaaatgg caatggctgc
     1681 cctgctaata ttgagtcgcg ttgtttatta caaggtaatg gtgatttaaa acctgaaaca
     1741 tcggtaaaca aagagctcgg tattcagttc caaaaagata tcgtgaatgc gagcttaact
     1801 tggttccgta atgattataa agataagatt gttgcgggta ctcatgttgt tggaacagtt
     1861 gatggctcaa gtacaaatgc aaatacagga gctgtgacca atacgaagtg gaatattttg
     1921 cgttgggaaa atacgcctaa agccttaatt caaggttttg aaggaagttt gggattagat
     1981 ttcggtgata ttcgctggac taataacttt acctacatga tggactcgaa agacaagcaa
     2041 actgggaatc cattatcttt agttccaatc tatacaatta actcaatttt tgattatgac
     2101 attactgatc aattggatgt aaattttgta tttactcaat atggtcgtca aaaatcacgt
     2161 caatttgcag agaatagact tgaatccggt ataggttcag gaggtgcgaa ttctgcgctt
     2221 aagccaagta cggtaaaaag ttatagtacc gctggtatta atgttggtta taagttttca
     2281 gaccaaatta gtacgcgtgt tggtgtgagt aatctgtttg ataaacaaat tttaagagac
     2341 agcaattcta ttagccaaac ttataatgag ccaggtcgag cttattacgc atctttaaaa
     2401 tattcttttt aaggatccga attcgagctc cgtcgacaag cttgcggccg cactcgagca
     2461 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     2521 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     2581 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     2641 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2701 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2761 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2821 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2881 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2941 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3001 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3061 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3121 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3181 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3241 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3301 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3361 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3421 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3481 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3541 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3601 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3661 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3721 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     3781 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3841 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3901 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3961 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4021 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4081 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4141 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4201 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4261 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4321 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4381 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4441 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4501 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4561 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4621 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4681 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4741 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4801 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4861 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4921 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4981 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     5041 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     5101 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     5161 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5221 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5281 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     5341 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     5401 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     5461 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     5521 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     5581 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     5641 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     5701 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     5761 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     5821 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     5881 gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg
     5941 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
     6001 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
     6061 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
     6121 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
     6181 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     6241 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6301 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     6361 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     6421 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     6481 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     6541 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     6601 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     6661 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     6721 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     6781 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     6841 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     6901 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     6961 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     7021 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     7081 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     7141 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     7201 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     7261 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     7321 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     7381 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     7441 tatgcaactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     7501 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     7561 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     7621 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     7681 ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac
     7741 tatagg
//
