LOCUS       sequence-232481-        6709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  precursor outer membrane pore protein E, phoE with mutations Alanine
            249 to Cysteine and Glutamine 307 to Cysteine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6709)
  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 6709)
  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: pAL950
FEATURES             Location/Qualifiers
     source          1..6709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..27
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     111..130
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        111..129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    130..154
                     /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             169..191
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1316..1334)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1330..1377
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1611..1629)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1697..1801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1802..1870,1871..2662)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRMHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2020..2039)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2833..3421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3322..3341
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3575..3592
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3607..3746
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3672..3691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3832..3854
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3848..4039)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5351..6433)
                     /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
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSVRLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6395..6414
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6434..6511)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
       61 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact
      121 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
      181 taagaaggag atatacatat gaaaaagagc actctggcat tagtggtgat gggcattgtg
      241 gcatctgcat ctgtacaggc tgcagaaata tataataaag acggtaataa actggatgtc
      301 tatggcaaag ttaaagccat gcattatatg agtgataacg ccagtaaaga tggcgaccag
      361 agttatatcc gttttggttt caaaggcgaa acacaaatta acgatcaact gactggttat
      421 ggtcgttggg aagcagagtt tgccggtaat aaagcagaga gtgatactgc acagcaaaaa
      481 acgcgtctcg cttttgccgg gttgaaatat aaagatttgg gttctttcga ttatggtcgt
      541 aacctggggg cgttgtatga cgtggaagcc tggaccgata tgttcccgga atttggtggc
      601 gattcctcgg cgcagaccga caactttatg accaaacgcg ccagcggtct ggcgacgtat
      661 cggaacaccg acttcttcgg cgttatcgat ggcctgaact taaccctgca atatcaaggg
      721 aaaaacgaaa accgcgacgt taaaaagcaa aacggcgatg gcttcggcac gtcattgacg
      781 tacgactttg gcggcagcga tttcgccatt agtggggcct ataccaactc agatcgcacc
      841 aacgagcaga acctgcaaag ccgtggcaca ggcaagcgtg cagaagcatg ggcaacaggt
      901 ctgaaatacg atgccaataa tatttatctg gcaactttct attctgaaac acgcaaaatg
      961 acgccaataa ctggcggctt tgccaataag acacagaact ttgaatgtgt cgctcaatac
     1021 cagtttgact ttggtctgcg tccatcgctg ggttatgtct tatcgaaagg gaaagatatt
     1081 gaaggtatcg gtgatgaaga tctggtcaat tatatcgacg tcggtgctac gtattatttc
     1141 aacaaaaata tgtcagcgtt tgttgattat aaaatcaact gtctggatag cgataacaaa
     1201 ttgaatatta ataatgatga tattgtcgcg gttggcatga cgtatcagtt ttaatgaata
     1261 ttgccggatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     1321 gagcaataac tagcataacc ccttggggcc tctaaacggg tctttagggg ttttttgctg
     1381 aaaggaggaa ctatatccgg atatcccgca agaggcccgg cagtaccggc ataaccaagc
     1441 ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt
     1501 tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
     1561 agcttatcga tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat
     1621 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     1681 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1741 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1801 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1861 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1921 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1981 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2041 ccgtgttgac gccgggcaag agcaactcgg tcgccgcatg cactattctc agaatgactt
     2101 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2161 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2221 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2281 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2341 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2401 ttcccgacaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2461 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2521 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2581 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2641 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2701 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2761 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2821 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2881 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2941 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3001 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3061 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3121 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3181 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3241 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3301 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3361 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3421 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3481 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3541 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3601 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     3661 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     3721 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     3781 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3841 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     3901 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3961 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4021 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     4081 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     4141 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     4201 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     4261 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     4321 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     4381 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     4441 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     4501 cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg
     4561 acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt
     4621 gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca
     4681 ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag
     4741 ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc
     4801 cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg
     4861 aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat
     4921 cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc
     4981 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt
     5041 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     5101 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     5161 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     5221 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     5281 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     5341 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5401 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     5461 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     5521 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     5581 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     5641 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     5701 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     5761 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     5821 gatatttatg ccagccagcc agacgcagac gcaccgagac agaacttaat gggcccgcta
     5881 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     5941 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     6001 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     6061 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     6121 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     6181 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     6241 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     6301 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     6361 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     6421 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     6481 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     6541 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     6601 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     6661 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatctt
//
