LOCUS       sequence-218609-        7763 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses PafA-XIAP in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7763)
  AUTHORS   Liu Q, Zheng J, Sun W, Huo Y, Zhang L, Hao P, Wang H, Zhuang M
  TITLE     A proximity-tagging system to identify membrane protein-protein 
            interactions.
  JOURNAL   Nat Methods. 2018 Aug 13. pii: 10.1038/s41592-018-0100-5. doi: 
            10.1038/s41592-018-0100-5.
  PUBMED    30104635
REFERENCE   2  (bases 1 to 7763)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2018 Aug 13. pii: 10.1038/s41592-018-0100-5. doi: 
            10.1038/s41592-018-0100-5."
COMMENT     Sequence Label: pGEX6p-1-PafA-XIAP
FEATURES             Location/Qualifiers
     source          1..7763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..30
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2902..2924)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3069..3087)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3155..3259
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3260..3328,3329..4120)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3478..3497)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4291..4879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4780..4799
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5033..5050
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5123..5200
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5201..6283
                     /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(5220..5239)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6332..6337,6338..6355,6356..6362)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6370..6386
                     /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     6394..6410
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6394..6410
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6406..6933
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6426..6443)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6426..6442)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6435..6457)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(7035..7040,7041..7056,7057..7063)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    7071..7087
                     /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)."
     CDS             7110..7763
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     7721..7743
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
ORIGIN
        1 tcggatctgg aagttctgtt ccaggggccc ctgggatccc cggaattccc gggtcgacga
       61 ctcgagatga gtaccgtgga atccgcattg acccgcagga tcatgggcat tgaaacggag
      121 tatggcctca cctttgttga tggtgattcc aaaaagcttc gcccagatga gatagctcga
      181 aggatgtttc gtcccatcgt ggagaaatat tccagctcta atatcttcat acccaatggt
      241 tcccggttgt atcttgatgt gggttcccac ccggagtacg ccaccgccga gtgtgataat
      301 ttgacccagc tgatcaattt tgaaaaagct ggcgatgtta ttgcagatcg catggctgta
      361 gatgccgaag agtcgctggc gaaagaagac attgctgggc aggtgtacct gtttaaaaac
      421 aatgtcgatt ccgtgggcaa ttcttatggc tgccacgaaa actaccttgt gggtcgctcc
      481 atgccgttga aggcgttggg taaaaggctg atgccgtttc tgattacccg ccagctcatc
      541 tgcggcgccg gcaggatcca tcaccccaat cctttggata aaggcgaatc cttccccttg
      601 ggctactgca tatcccagcg ctctgaccac gtgtgggagg gcgtatcaag tgccaccact
      661 agatcacgcc ccattatcaa cacccgtgat gagccacatg cggattccca ttcttaccgc
      721 aggctgcacg tgattgtggg tgatgccaac atggcagagc ccagcatcgc gttgaaggtc
      781 ggctccacgt tgctggttct ggaaatgatt gaggcagatt tcggtttgcc cagcttagag
      841 cttgccaatg atattgcctc aattagggaa atctcccgcg atgcaacagg atccacactg
      901 ttgtccctga aagatggcac caccatgact gccttgcaga tccagcaggt ggtctttgag
      961 catgcctcga agtggttgga gcagcgcccc gaaccagaat tttctggcac ctccaacaca
     1021 gagatggccc gcgtgctgga tctgtggggt cgcatgttga aagcgattga gtccggtgat
     1081 ttcagcgaag tggatacaga aattgactgg gtgatcaaaa agaagctcat tgatcgtttc
     1141 attcagcgcg gcaaccttgg gttggatgat ccaaaacttg cccaagtgga cttgacttat
     1201 cacgatatta ggccaggtag aggcctattt agcgtgctgc aaagccgcgg catgatcaaa
     1261 cggtggacta ctgatgaggc gattttagct gcggtggata ccgctcctga tacaacacgt
     1321 gctcatttgc gcgggcgaat ccttaaagcg gcggatactc tgggagtacc tgtgactgtc
     1381 gattggatgc gtcacaaggt caaccgaccg gagccacaat cggtggaatt gggggatcct
     1441 ttttccgcag tgaattccga ggttgatcag ttgattgagt atatgaccgt tcatgcggag
     1501 agctatcgca gtggcggttc tggtggcggt tctggtatga cttttaacag ttttgaagga
     1561 tctaaaactt gtgtacctgc agacatcaat aaggaagaag aatttgtaga agagtttaat
     1621 agattaaaaa cttttgctaa ttttccaagt ggtagtcctg tttcagcatc aacactggca
     1681 cgagcagggt ttctttatac tggtgaagga gataccgtgc ggtgctttag ttgtcatgca
     1741 gctgtagata gatggcaata tggagactca gcagttggaa gacacaggaa agtatcccca
     1801 aattgcagat ttatcaacgg cttttatctt gaaaatagtg ccacgcagtc tacaaattct
     1861 ggtatccaga atggtcagta caaagttgaa aactatctgg gaagcagaga tcattttgcc
     1921 ttagacaggc catctgagac acatgcagac tatcttttga gaactgggca ggttgtagat
     1981 atatcagaca ccatataccc gaggaaccct gccatgtata gtgaagaagc tagattaaag
     2041 tcctttcaga actggccaga ctatgctcac ctaaccccaa gagagttagc aagtgctgga
     2101 ctctactaca caggtattgg tgaccaagtg cagtgctttt gttgtggtgg aaaactgaaa
     2161 aattgggaac cttgtgatcg tgcctggtca gaacacaggc gacactttcc taattgcttc
     2221 tttgttttgg gccggaatct taatattcga agtgaatctg atgctgtgag ttctgatagg
     2281 aatttcccaa attcaacaaa tcttccaaga aatccatcca tggcagatta tgaagcacgg
     2341 atctttactt ttgggacatg gatatactca gttaacaagg agcagcttgc aagagctgga
     2401 ttttatgctt taggtgaagg tgataaagta aagtgctttc actgtggagg agggctaact
     2461 gattggaagc ccagtgaaga cccttgggaa caacatgcta aatggtatcc agggtgcaaa
     2521 tatctgttag aacagaaggg acaagaatat ataaacaata ttcatttaac tcattcactt
     2581 gaggagtgtc tggtaagaac tactgagaaa acaccatcac taactagaag aattgatgat
     2641 accatcttcc aaaatcctat ggtacaagaa gctatacgaa tggggttcag tttcaaggac
     2701 attaagaaaa taatggagga aaaaattcag atatctggga gcaactataa atcacttgag
     2761 gttctggttg cagatctagt gaatgctcag aaagacagta tgcaagatga gtcaagtcag
     2821 acttcattac agaaagagta agcggccgca tcgtgactga ctgacgatct gcctcgcgcg
     2881 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2941 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3001 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag
     3061 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3121 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3181 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3241 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3301 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3361 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3421 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3481 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     3541 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3601 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3661 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3721 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3781 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3841 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3901 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3961 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4021 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4081 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4141 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4201 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4261 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4321 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4381 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4441 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4501 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4561 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4621 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4681 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4741 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4801 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4861 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4921 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4981 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5041 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     5101 tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc
     5161 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     5221 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     5281 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     5341 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     5401 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     5461 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     5521 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     5581 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     5641 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     5701 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     5761 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     5821 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     5881 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     5941 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     6001 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     6061 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     6121 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     6181 aaccacgcta gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6241 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6301 taagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6361 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     6421 gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     6481 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     6541 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg
     6601 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc
     6661 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc
     6721 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg
     6781 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat
     6841 ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca
     6901 tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag
     6961 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat
     7021 tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg
     7081 ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat tggaaaatta
     7141 agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa tatgaagagc
     7201 atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg
     7261 agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag tctatggcca
     7321 tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag
     7381 agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg agaattgcat
     7441 atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct gaaatgctga
     7501 aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat gtaacccatc
     7561 ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca atgtgcctgg
     7621 atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca caaattgata
     7681 agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa gccacgtttg
     7741 gtggtggcga ccatcctcca aaa
//
