LOCUS       sequence-131124-        6805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pertussis Toxin Subunit 1 Yeast Display Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6805)
  AUTHORS   Kowalsky CA, Faber MS, Nath A, Dann HE, Kelly VW, Liu L, Shanker P, 
            Wagner EK, Maynard JA, Chan C, Whitehead TA
  TITLE     Rapid fine conformational epitope mapping using comprehensive 
            mutagenesis and deep sequencing.
  JOURNAL   J Biol Chem. 2015 Oct 30;290(44):26457-70. doi: 
            10.1074/jbc.M115.676635. Epub 2015 Aug 20.
  PUBMED    26296891
REFERENCE   2  (bases 1 to 6805)
  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: 
            "10.1074/jbc.M115.676635"; journalName: "J Biol Chem"; date: 
            "2015-10-30- 30"; volume: "290"; issue: "44"; pages: "26457-70"
COMMENT     Sequence Label: pETCON-PTX-S1-220
FEATURES             Location/Qualifiers
     source          1..6805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..18
                     /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     7..29
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     26..42
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     26..42
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        63..81
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        111..763
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    315..432
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     727..750
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1082..1093
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1094..1120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1868..1897
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2226..2245)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2227..2245)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2252..2269)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2252..2268)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2261..2283)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2409..2864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2496..2515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2706..2727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2967..3641)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3642..3922)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3943..3962)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4062..4084
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4122..4140)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4181..4684
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4721..4825
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4826..4894,4895..5686)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5044..5063)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5857..6445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6346..6365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6599..6616
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6733..6754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6769..6774,6775..6792,6793..6799)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
       61 gaaattaacc ctcactaaag ggaacaaaag ctggtaccaa ttccttgaat tttcaaaaat
      121 tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc
      181 accatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat
      241 tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt
      301 gctatattga agtacggatt agaagccgcc gagcgggtga cagccctccg aaggaagact
      361 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
      421 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
      481 aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag
      541 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
      601 atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt taactaatac
      661 tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa
      721 attgttaata tacctctata ctttaacgtc aaggagaaaa aaccccggat cgaattccct
      781 acttcataca ttttcaatta agatgcagtt acttcgctgt ttttcaatat tttctgttat
      841 tgcttcagtt ttagcacagg aactgacaac tatatgcgag caaatcccct caccaacttt
      901 agaatcgacg ccgtactctt tgtcaacgac tactattttg gccaacggga aggcaatgca
      961 aggagttttt gaatattaca aatcagtaac gtttgtcagt aattgcggtt ctcacccctc
     1021 aacaactagc aaaggcagcc ccataaacac acagtatgtt tttaaggaca atagctcgac
     1081 gattgaaggt agatacccat acgacgttcc agactacgct ctgcaggcta gtggtggagg
     1141 aggctctggt ggaggcggta gcggaggcgg agggtcggct agccatatgg acgatcctcc
     1201 cgccaccgta taccgctatg actcccgccc gccggaggac gttttccaga acggattcac
     1261 ggcgtgggga aacaacgaca atgtgctcga ccatctgacc ggacgttcct gccaggtcgg
     1321 cagcagcaac agcgctttcg tctccaccag cagcagccgg cgctataccg aggtctatct
     1381 cgaacatcgc atgcaggaag cggtcgaggc cgaacgcgcc ggcaggggca ccggccactt
     1441 catcggctac atctacgaag tccgcgccga caacaatttc tacggcgccg ccagctcgta
     1501 cttcgaatac gtcgacactt atggcgacaa tgccggccgt atcctcgccg gcgcgctggc
     1561 cacctaccag agcgaatatc tggcacaccg gcgcattccg cccgaaaaca tccgcagggt
     1621 aacgcgggtc tatcacaacg gcatcaccgg cgagaccacg accacggagt attccaacgc
     1681 tcgctacgtc agccagcaga ctcgcgccaa tcccaacccc tacacatcgc gaaggtccgt
     1741 agcgtcgatc gtcggcacat tggtgcgcat ggcgccggtg ataggcgctt gcatggcgcg
     1801 gcaggccgaa agctccgagg ccatggcagc ctggtccgaa cgcgccggcc tcgagggggg
     1861 cggatccgaa caaaagctta tttctgaaga ggacttgtaa tagagatctg ataacaacag
     1921 tgtagatgta acaaaatcga ctttgttccc actgtacttt tagctcgtac aaaatacaat
     1981 atacttttca tttctccgta aacaacatgt tttcccatgt aatatccttt tctatttttc
     2041 gttccgttac caactttaca catactttat atagctattc acttctatac actaaaaaac
     2101 taagacaatt ttaattttgc tgcctgccat atttcaattt gttataaatt cctataattt
     2161 atcctattag tagctaaaaa aagatgaatg tgaatcgaat cctaagagaa ttgagctcca
     2221 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     2281 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2341 ggcgtaatag cgaagaggcc cgcaccgatc gcctttccca acagttgcgc agcctgaatg
     2401 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2461 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2521 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2581 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2641 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2701 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2761 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2821 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     2881 ctccttacgc atctgtgcgg tatttcacac cgcatagatc ggcaagtgca caaacaatac
     2941 ttaaataaat actactcagt aataacctat ttcttagcat ttttgacgaa atttgctatt
     3001 ttgttagagt cttttacacc atttgtctcc acacctccgc ttacatcaac accaataacg
     3061 ccatttaatc taagcgcatc accaacattt tctggcgtca gtccaccagc taacataaaa
     3121 tgtaagcttt cggggctctc ttgccttcca acccagtcag aaatcgagtt ccaatccaaa
     3181 agttcacctg tcccacctgc ttctgaatca aacaagggaa taaacgaatg aggtttctgt
     3241 gaagctgcac tgagtagtat gttgcagtct tttggaaata cgagtctttt aataactggc
     3301 aaaccgagga actcttggta ttcttgccac gactcatctc catgcagttg gacgatatca
     3361 atgccgtaat cattgaccag agccaaaaca tcctccttag gttgattacg aaacacgcca
     3421 accaagtatt tcggagtgcc tgaactattt ttatatgctt ttacaagact tgaaattttc
     3481 cttgcaataa ccgggtcaat tgttctcttt ctattgggca cacatataat acccagcaag
     3541 tcagcatcgg aatctagagc acattctgcg gcctctgtgc tctgcaagcc gcaaactttc
     3601 accaatggac cagaactacc tgtgaaatta ataacagaca tactccaagc tgcctttgtg
     3661 tgcttaatca cgtatactca cgtgctcaat agtcaccaat gccctccctc ttggccctct
     3721 ccttttcttt tttcgaccga attaattctt aatcggcaaa aaaagaaaag ctccggatca
     3781 agattgtacg taaggtgaca agctattttt caataaagaa tatcttccac tactgccatc
     3841 tggcgtcata actgcaaagt acacatatat tacgatgctg ttctattaaa tgcttcctat
     3901 attatatata tagtaatgtc gtgatctatg gtgcactctc agtacaatct gctctgatgc
     3961 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4021 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4081 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4141 tttataggtt aatgtcatga taataatggt ttcttagacg gatcgcttgc ctgtaactta
     4201 cacgcgcctc gtatctttta atgatggaat aatttgggaa tttactctgt gtttatttat
     4261 ttttatgttt tgtatttgga ttttagaaag taaataaaga aggtagaaga gttacggaat
     4321 gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac tttacatata
     4381 tatttattag acaagaaaag cagattaaat agatatacat tcgattaacg ataagtaaaa
     4441 tgtaaaatca caggattttc gtgtgtggtc ttctacacag acaagatgaa acaattcggc
     4501 attaatacct gagagcagga agagcaagat aaaaggtagt atttgttggc gatcccccta
     4561 gagtctttta catcttcgga aaacaaaaac tattttttct ttaatttctt tttttacttt
     4621 ctatttttaa tttatatatt tatattaaaa aatttaaatt ataattattt ttatagcacg
     4681 tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4741 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4801 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4861 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4921 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4981 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5041 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5101 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5161 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5221 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5281 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5341 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5401 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5461 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5521 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5581 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5641 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5701 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5761 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5821 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5881 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5941 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6001 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6061 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6121 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6181 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6241 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6301 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6361 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6421 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6481 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6541 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6601 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6661 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6721 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6781 gcttccggct cgtatgttgt gtgga
//
