LOCUS       sequence-232053-        5342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  precusor outer membrane protein A, ompA, with mutations Cysteine 290
            Serine, Glycine 244 Cysteine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5342)
  AUTHORS   Bariya P, Randall LL
  TITLE     Co-assembly of SecYEG and SecA fully restores the properties of the 
            native translocon.
  JOURNAL   J Bacteriol. 2018 Oct 1. pii: JB.00493-18. doi: 10.1128/JB.00493-18.
  PUBMED    30275279
REFERENCE   2  (bases 1 to 5342)
  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: "J 
            Bacteriol. 2018 Oct 1. pii: JB.00493-18. doi: 10.1128/JB.00493-18."
COMMENT     Sequence Label: pAL612
FEATURES             Location/Qualifiers
     source          1..5342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(715..720,721..737,738..744)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    752..768
                     /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     757..779
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     sig_peptide     945..1007
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     primer_bind     complement(2048..2065)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2218..2304
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2396..2423
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2442..2533
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2534..2602,2603..3394)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2752..2771)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3565..4153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4054..4073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4307..4324
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4339..4479
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4405..4424)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4565..4587
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4665..4742
                     /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             join(4743..5342,1..483)
                     /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(4762..4781)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
       61 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
      121 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
      181 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
      241 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
      301 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
      361 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
      421 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      481 tgagcgcaac gcaattaatg taagttagcg cgaattgatc tggtttgaca gcttatcatc
      541 gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct
      601 gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
      661 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca
      721 attaatcatc cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga
      781 aacagaccat ggaattccca gactttacat cgccaggggt gctcggcata agccgaagat
      841 atcggtagag ttaatattga gcagatcccc cggtgaagga tttaaccgtg ttatctcgtt
      901 ggagatattc atggcgtatt ttggatgata acgaggcgca aaaaatgaaa aagacagcta
      961 tcgcgattgc agtggcactg gctggtttcg ctaccgtagc gcaggccgct ccgaaagata
     1021 acacctggta cactggtgct aaactgggct ggtcccagta ccatgacact ggtttcatca
     1081 acaacaatgg cccgacccat gaaaaccaac tgggcgctgg tgcttttggt ggttaccagg
     1141 ttaacccgta tgttggcttt gaaatgggtt acgactggtt aggtcgtatg ccgtacaaag
     1201 gcagcgttga aaacggtgca tacaaagctc agggcgttca actgaccgct aaactgggtt
     1261 acccaatcac tgacgacctg gacatctaca ctcgtctggg tggcatggta tggcgtgcag
     1321 acactaaatc caacgtttat ggtaaaaacc acgacaccgg cgtttctccg gtcttcgctg
     1381 gcggtgttga gtacgcgatc actcctgaaa tcgctacccg tctggaatac cagtggacca
     1441 acaacatcgg tgacgcacac accatcggca ctcgtccgga caacggcatg ctgagcctgg
     1501 gtgtttccta ccgtttcggt cagggcgaag cagctccagt agttgctccg gctccagctc
     1561 cggcaccgga agtacagacc aagcacttca ctctgaagtc tgacgttctg ttcaacttca
     1621 acaaagcaac cctgaaaccg gaaggtcagg ctgctctgga tcagctgtac agccagctga
     1681 gcaacctgga tccgaaagac ggttccgtag ttgttctggg ttacaccgac cgcatctgtt
     1741 ctgacgctta caaccagggt ctgtccgagc gccgtgctca gtctgttgtt gattacctga
     1801 tctccaaagg tatcccggca gacaagatct ccgcacgtgg tatgggcgaa tccaacccgg
     1861 ttactggcaa cacctctgac aacgtgaaac agcgtgctgc actgatcgac tgcctggctc
     1921 cggatcgtcg cgtagagatc gaagttaaag gtatcaaaga cgttgtaact cagccgcagg
     1981 cttaatctag agtcgacctg caggcatgca agcttggctg ttttggcgga tgagagaaga
     2041 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     2101 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     2161 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     2221 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2281 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     2341 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     2401 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     2461 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2521 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2581 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2641 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2701 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2761 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     2821 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2881 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2941 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3001 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3061 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3121 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3181 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3241 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3301 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3361 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3421 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3481 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3541 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3601 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3661 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3721 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3781 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3841 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3901 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3961 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4021 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4081 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4141 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4201 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4261 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4321 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4381 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4441 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4501 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4561 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4621 cagatcaatt cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc atcgaatggt
     4681 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     4741 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     4801 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     4861 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     4921 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     4981 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     5041 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     5101 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     5161 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     5221 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     5281 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     5341 gc
//
