LOCUS       sequence-59600-e        7069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7069)
  TITLE     pETCON Yeast Surface Display Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7069)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pETcon(I-LtrI)
FEATURES             Location/Qualifiers
     source          1..7069
                     /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             2132..2161
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2490..2509)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2491..2509)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2516..2533)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2516..2532)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2525..2547)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2673..3128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2760..2779)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2970..2991
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3231..3905)
                     /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(3906..4186)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4207..4226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4326..4348
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4386..4404)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4445..4948
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4985..5089
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5090..5158,5159..5950)
                     /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(5308..5327)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6121..6709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6610..6629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6863..6880
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6997..7018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7033..7038,7039..7056,7057..7063)
                     /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 agccatatgt tcccagttca
     1201 agctagaaac gacaacatct ctccatggac tatcactggt ttcgctgacg ctgaatcttc
     1261 tttcatgttg actgtttcta aggactctaa gagaaacact ggttggtctg ttagaccaag
     1321 attcagaatc ggtttgcaca acaaggacgt gactatcttg aagtctatca gagaatactt
     1381 gggcgccggt atcatcactt ctgacaagga cgctagaatc agattcgaat ctttgaagga
     1441 attggaagtt gttatcaacc acttcgacaa gtacccattg atcactcaaa agagagctga
     1501 ctacttgttg ttcaagaagg ctttctactt aattaagaac aaggaacact tgactgaaga
     1561 aggtttgaac caaatcttga ctttgaaggc ttctttgaac ttgggtttgt ctgaagaatt
     1621 gaaggaagca ttcccaaaca ctatcccagc tgaaaagtta ctagttactg gtcaagaaat
     1681 cccagactct aactgggttg ctggtttcac tgctggtgaa ggttctttct acatcagaat
     1741 cgctaagaac tctactttga agactggtta ccaagttcaa tctgttttcc aaatcactca
     1801 agacacgcgt gacatcgaat tgatgaagaa cttgatctct tacttgaact gtggtaacat
     1861 cagaatcaga aagtacaagg gttctgaagg tatccacgac acttgtgttg acttggttgt
     1921 tactaacttg aacgacatca aggaaaagat catcccattc ttcaacaaga accacatcat
     1981 cggtgttaag ttgcaagact acagagactg gtgtaaggtt gttactttga tcgacaacaa
     2041 ggaacacttg acttctgaag gtttggaaaa gatccaaaag atcaaggaag gtatgaacag
     2101 aggtagatct ttgctcgagg ggggcggatc cgaacaaaag cttatttctg aagaggactt
     2161 gtaatagaga tctgataaca acagtgtaga tgtaacaaaa tcgactttgt tcccactgta
     2221 cttttagctc gtacaaaata caatatactt ttcatttctc cgtaaacaac atgttttccc
     2281 atgtaatatc cttttctatt tttcgttccg ttaccaactt tacacatact ttatatagct
     2341 attcacttct atacactaaa aaactaagac aattttaatt ttgctgcctg ccatatttca
     2401 atttgttata aattcctata atttatccta ttagtagcta aaaaaagatg aatgtgaatc
     2461 gaatcctaag agaattgagc tccaattcgc cctatagtga gtcgtattac aattcactgg
     2521 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     2581 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccttt
     2641 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     2701 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2761 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2821 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2881 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2941 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3001 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3061 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     3121 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3181 gatcggcaag tgcacaaaca atacttaaat aaatactact cagtaataac ctatttctta
     3241 gcatttttga cgaaatttgc tattttgtta gagtctttta caccatttgt ctccacacct
     3301 ccgcttacat caacaccaat aacgccattt aatctaagcg catcaccaac attttctggc
     3361 gtcagtccac cagctaacat aaaatgtaag ctttcggggc tctcttgcct tccaacccag
     3421 tcagaaatcg agttccaatc caaaagttca cctgtcccac ctgcttctga atcaaacaag
     3481 ggaataaacg aatgaggttt ctgtgaagct gcactgagta gtatgttgca gtcttttgga
     3541 aatacgagtc ttttaataac tggcaaaccg aggaactctt ggtattcttg ccacgactca
     3601 tctccatgca gttggacgat atcaatgccg taatcattga ccagagccaa aacatcctcc
     3661 ttaggttgat tacgaaacac gccaaccaag tatttcggag tgcctgaact atttttatat
     3721 gcttttacaa gacttgaaat tttccttgca ataaccgggt caattgttct ctttctattg
     3781 ggcacacata taatacccag caagtcagca tcggaatcta gagcacattc tgcggcctct
     3841 gtgctctgca agccgcaaac tttcaccaat ggaccagaac tacctgtgaa attaataaca
     3901 gacatactcc aagctgcctt tgtgtgctta atcacgtata ctcacgtgct caatagtcac
     3961 caatgccctc cctcttggcc ctctcctttt cttttttcga ccgaattaat tcttaatcgg
     4021 caaaaaaaga aaagctccgg atcaagattg tacgtaaggt gacaagctat ttttcaataa
     4081 agaatatctt ccactactgc catctggcgt cataactgca aagtacacat atattacgat
     4141 gctgttctat taaatgcttc ctatattata tatatagtaa tgtcgtgatc tatggtgcac
     4201 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4261 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4321 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4381 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4441 gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg
     4501 ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata
     4561 aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc
     4621 aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat
     4681 acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac
     4741 acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg
     4801 tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt
     4861 ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta
     4921 aattataatt atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa
     4981 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5041 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5101 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5161 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5221 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5281 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5341 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5401 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5461 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5521 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5581 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5641 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5701 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5761 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5821 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5881 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5941 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6001 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6061 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6121 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6181 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6241 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6301 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6361 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6421 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6481 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6541 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6601 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6661 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6721 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6781 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6841 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     6901 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     6961 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     7021 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtgga
//
