LOCUS       sequence-90375-e        4938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of APX W41F inducible by IPTG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4938)
  AUTHORS   Dwyer DJ, Belenky PA, Yang JH, MacDonald IC, Martell JD, Takahashi 
            N, Chan CT, Lobritz MA, Braff D, Schwarz EG, Ye JD, Pati M, 
            Vercruysse M, Ralifo PS, Allison KR, Khalil AS, Ting AY, Walker GC, 
            Collins JJ
  TITLE     Antibiotics induce redox-related physiological alterations as part 
            of their lethality.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 May 20;111(20):E2100-9. doi: 
            10.1073/pnas.1401876111. Epub 2014 May 6.
  PUBMED    24803433
REFERENCE   2  (bases 1 to 4938)
  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"; doi: 
            "10.1073/pnas.1401876111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-05-20- 20"; volume: "111"; issue: "20"; pages: "E2100-9"
COMMENT     Sequence Label: pTRC-APX(W41F) dimeric
FEATURES             Location/Qualifiers
     source          1..4938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     348..365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    380..520
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(446..465)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     606..628
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        706..783
                     /gene="lacI (mutant)"
                     /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             784..1866
                     /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     complement(803..822)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2098..2103,2104..2120,2121..2127)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2135..2151
                     /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)."
     primer_bind     2140..2162
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             2178..2195
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3027..3044)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3197..3283
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3375..3402
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3421..3512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3513..3581,3582..4373)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAWQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3731..3750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4544..4938,1..194)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
       61 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      301 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      361 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
      421 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
      481 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
      541 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
      601 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
      661 gcagatcaat tcgcgcgcga aggcgaagcg gcatgcattt acgttgacac catcgaatgg
      721 tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
      781 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
      841 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
      901 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
      961 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     1021 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     1081 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     1141 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     1201 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     1261 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     1321 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     1381 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     1441 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     1501 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     1561 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
     1621 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     1681 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     1741 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     1801 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1861 cagtgagcgc aacgcaatta atgtaagtta gcgcgaattg atctggtttg acagcttatc
     1921 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
     1981 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     2041 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
     2101 acaattaatc atccggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca
     2161 ggaaacagac catggcacac caccaccacc accacaagga tccaggaaag tcatacccaa
     2221 ccgtgagccc agattaccag aaggctattg agaaggcaaa aaggaaactg aggggattca
     2281 tcgcagaaaa gaaatgcgcc ccactgatcc tgcgactggc tttccacagc gcaggaacat
     2341 ttgactccaa gaccaaaaca ggcggaccct ttgggactat caagcaccag gctgaactgg
     2401 cacatggggc caacaatggc ctggacattg ctgtgagact gctggagcca atcaaagaac
     2461 agttccccat tgtctcctac gcagacttct accagctggc cggcgtggtc gctgtggaga
     2521 ttaccggcgg ccccgaggtc cctttccacc caggaaggga ggacaaacct gaaccccctc
     2581 cagaggggag actgccagat gctaccaagg gcagcgacca tctgcgggac gtgttcggaa
     2641 aagcaatggg gctgtctgac caggatatcg tggccctgag tggagggcac accattggcg
     2701 ccgctcataa ggaacgctca ggcttcgagg gaccctggac aagcaaccct ctgatcttcg
     2761 acaattccta ctttactgaa ctgctgaccg gcgagaagga tggactgctg cagctgccct
     2821 ctgacaaagc cctgctgaca gatagcgtgt tcaggcctct ggtcgaaaag tatgcagccg
     2881 acgaggatgt gttcttcgcc gactatgctg aggcacacct gaaactgtct gaactgggat
     2941 tcgccgaggc agggaattct taagatcctc tagagtcgac ctgcaggcaa gcttggctgt
     3001 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     3061 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     3121 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     3181 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     3241 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     3301 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     3361 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     3421 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3481 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3541 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3601 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3661 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3721 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgcctggcaa gagcaactcg
     3781 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3841 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3901 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3961 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4021 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4081 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4141 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4201 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4261 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4321 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4381 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4441 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4501 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4561 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4621 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4681 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4741 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4801 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4861 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4921 acctacagcg tgagctat
//
