LOCUS       sequence-191141-        9134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  titin in DNA-binding domain (BD) vector, used for yeast-two hybrid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9134)
  TITLE     Shiang Lab CMYA5 project plasmids (unpublished)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9134)
  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            "Unpublished"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: TTN/pAS2-1
FEATURES             Location/Qualifiers
     source          1..9134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..1344
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(965..1012)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     1385..1403
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1441..1463)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1563..1582
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1603..1884
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1885..2559
                     /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"
     rep_origin      complement(2662..3117)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2799..2820)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3011..3030
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3243..3265
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3257..3274
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3258..3274
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(3308..3555)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3521..3539
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        4764..5468
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5496..5936
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     terminator      6772..6959
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6827..6851)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6983..6999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6983..6999)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6996..7018)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7007..7023
                     /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)."
     promoter        complement(join(7031..7037,7038..7055,7056..7061))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7076..7097
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7214..7231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7385..7973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7465..7484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8144..8935,8936..9004))
                     /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     8767..8786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9005..9109)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
       61 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
      121 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
      181 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
      241 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
      301 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
      361 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
      421 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
      481 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
      541 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
      601 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
      661 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
      721 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
      781 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
      841 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
      901 ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg
      961 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact tcaaagcgtt
     1021 tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg cacatacagc tcactgttca
     1081 cgtcgcacct atatctgcgt gttgcctgta tatatatata catgagaaga acggcatagt
     1141 gcgtgtttat gcttaaatgc gtacttatat gcgtctattt atgtaggatg aaaggtagtc
     1201 tagtacctcc tgtgatatta tcccattcca tgcggggtat cgtatgcttc cttcagcact
     1261 accctttagc tgttctatat gctgccactc ctcaattgga ttagtctcat ccttcaatgc
     1321 tatcatttcc tttgatattg gatcatacta agaaaccatt attatcatga cattaaccta
     1381 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1441 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1501 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1561 tgcggcatca gagcagattg tactgagagt gcaccataga tcaacgacat tactatatat
     1621 ataatatagg aagcatttaa tagaacagca tcgtaatata tgtgtacttt gcagttatga
     1681 cgccagatgg cagtagtgga agatattctt tattgaaaaa tagcttgtca ccttacgtac
     1741 aatcttgatc cggagctttt ctttttttgc cgattaagaa ttaattcggt cgaaaaaaga
     1801 aaaggagagg gccaagaggg agggcattgg tgactattga gcacgtgagt atacgtgatt
     1861 aagcacacaa aggcagcttg gagtatgtct gttattaatt tcacaggtag ttctggtcca
     1921 ttggtgaaag tttgcggctt gcagagcaca gaggccgcag aatgtgctct agattccgat
     1981 gctgacttgc tgggtattat atgtgtgccc aatagaaaga gaacaattga cccggttatt
     2041 gcaaggaaaa tttcaagtct tgtaaaagca tataaaaata gttcaggcac tccgaaatac
     2101 ttggttggcg tgtttcgtaa tcaacctaag gaggatgttt tggctctggt caatgattac
     2161 ggcattgata tcgtccaact gcatggagat gagtcgtggc aagaatacca agagttcctc
     2221 ggtttgccag ttattaaaag actcgtattt ccaaaagact gcaacatact actcagtgca
     2281 gcttcacaga aacctcattc gtttattccc ttgtttgatt cagaagcagg tgggacaggt
     2341 gaacttttgg attggaactc gatttctgac tgggttggaa ggcaagagag ccccgaaagc
     2401 ttacatttta tgttagctgg tggactgacg ccagaaaatg ttggtgatgc gcttagatta
     2461 aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga caaatggtgt aaaagactct
     2521 aacaaaatag caaatttcgt caaaaatgct aagaaatagg ttattactga gtagtattta
     2581 tttaagtatt gtttgtgcac ttgccgatct atgcggtgtg aaataccgca cagatgcgta
     2641 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2701 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2761 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2821 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2881 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2941 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3001 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3061 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     3121 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     3181 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     3241 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctttcgcgag ctcgagatcc
     3301 cgagcttgca aattaaagcc ttcgagcgtc ccaaaacctt ctcaagcaag gttttcagta
     3361 taatgttaca tgcgtacacg cgtctgtaca gaaaaaaaag aaaaatttga aatataaata
     3421 acgttcttaa tactaacata actataaaaa aataaatagg gacctagact tcaggttgtc
     3481 taactccttc cttttcggtt agagcggatg tggggggagg gcgtgaatgt aagcgtgaca
     3541 taactaatta catgatatcg acaaaggaaa aggggcctgt ttactcacag gcttttttca
     3601 agtaggtaat taagtcgttt ctgtcttttt ccttcttcaa cccaccaaag gccatcttgg
     3661 tacttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt
     3721 tttttttttt tttttttttt tttttttttt tttttttttg atagaaataa tacagaagta
     3781 gatgttgaat tagattaaac tgaagatata taatttattg gaaaatacat agagtttttg
     3841 ttgatgcgct taagcgatca attcaacaac accaccagca gctctgattt tttcttcagc
     3901 caacttggag acgaatctag ctttgacgat aactggaaca tttgggattc tacccttacc
     3961 caagatctta ccgtaaccgg ctgccaaagt gtcaataact ggagcagttt ccttagaagc
     4021 agatttcaag tattggtctc tcttgtcttc tgggatcaat gtccacaatt tgtccaagtt
     4081 caagactggc ttccagaaat gagcttgttg cttgtggaag tatctcatac caaccttacc
     4141 gaaataacct ggatggtatt tatccatgtt aattctgtgg tgatgttgac caccggccat
     4201 acctctacca ccggggtgct ttctgtgctt accgatacga cctttaccgg ctgagacgtg
     4261 acctctgtgc tttctagtct tagtgaatct ggaaggcatt cttgattagt tggatgattg
     4321 ttctgggatt taatgcaaaa aaatcactaa gaaggaaaaa aatcaacgga gaaagcaaac
     4381 gccatcttaa atatacggga tacagatgaa aggtttgaac ctatctggga aaatacgcat
     4441 taaacaagcg aaaaactgcg aggaaaattg tttgcgtctc tgcgggctat tcacgcgcca
     4501 gaggaaaaat aggaaaaata acaggcatta gaaaaataat tttgattttg gtaatgtgtg
     4561 ggtccctggt gtacagatgt tacattggtt acagtactct tgtttttgct gtgtttttcg
     4621 atgaatctcc aaaatggttg ttagcacatg gaagagtcac cgatgctaag ttatctctat
     4681 gtaagctacg tggcgtgact tttgatgaag ccgcacaaga gatacaggat tggcaactgc
     4741 aaatagaatc tggggatcta gatatccttt tgttgtttcc gggtgtacaa tatggacttc
     4801 ctcttttctg gcaaccaaac ccatacatcg ggattcctat aataccttcg ttggtctccc
     4861 taacatgtag gtggcggagg ggagatatac aatagaacag ataccagaca agacataatg
     4921 ggctaaacaa gactacacca attacactgc ctcattgatg gtggtacata acgaactaat
     4981 actgtagccc tagacttgat agccatcatc atatcgaagt ttcactaccc tttttccatt
     5041 tgccatctat tgaagtaata ataggcgcat gcaacttctt ttcttttttt ttcttttctc
     5101 tctcccccgt tgttgtctca ccatatccgc aatgacaaaa aaatgatgga agacactaaa
     5161 ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt tgttccagag ctgatgaggg
     5221 gtatctcgaa gcacacgaaa ctttttcctt ccttcattca cgcacactac tctctaatga
     5281 gcaacggtat acggccttcc ttccagttac ttgaatttga aataaaaaaa agtttgctgt
     5341 cttgctatca agtataaata gacctgcaat tattaatctt ttgtttcctc gtcattgttc
     5401 tcgttccctt tcttccttgt ttctttttct gcacaatatt tcaagctata ccaagcatac
     5461 aatcaactcc aagcttgaag caagcctcct gaaagatgaa gctactgtct tctatcgaac
     5521 aagcatgcga tatttgccga cttaaaaagc tcaagtgctc caaagaaaaa ccgaagtgcg
     5581 ccaagtgtct gaagaacaac tgggagtgtc gctactctcc caaaaccaaa aggtctccgc
     5641 tgactagggc acatctgaca gaagtggaat caaggctaga aagactggaa cagctatttc
     5701 tactgatttt tcctcgagaa gaccttgaca tgattttgaa aatggattct ttacaggata
     5761 taaaagcatt gttaacagga ttatttgtac aagataatgt gaataaagat gccgtcacag
     5821 atagattggc ttcagtggag actgatatgc ctctaacatt gagacagcat agaataagtg
     5881 cgacatcatc atcggaagag agtagtaaca aaggtcaaag acagttgact gtatcgccgg
     5941 tattgcaata cccagctttg actcatatgg ccatgggctc cagaaatgta tactcccttg
     6001 aaatccgaaa tgcatcagtc agcgacagtg gaaagtacac aattaaggcc aaaaatttcc
     6061 gtggccagtg ttcagctaca gcttccttaa tggtccttcc tctagttgaa gaaccttcca
     6121 gagaggtagt attgagaaca agtggtgaca caagcttgca aggaagcttc tcgtctcagt
     6181 cagtccaaat gtctgcctcc aagcaggagg cctccttcag cagtttcagc agcagcagtg
     6241 ctagcagcat gactgagatg aaatttgcaa gcatgtctgc ccaaagcatg tcctccatgc
     6301 aagagtcctt tgtagaaatg agttccagca gctttatggg aatatctaat atgacacaac
     6361 tggaaagctc aactagtaaa atgcttaaag caggcataag aggaattccg cctaaaattg
     6421 aagctcttcc atctgatatc agcattgatg aaggcaaagt tctaacagta gcctgtgctt
     6481 tcacgggtga gcctacccca gaagtaacat ggtcctgtgg tggaagaaaa atccacagtc
     6541 aagaacaggg gaggttccac attgaaaaca cagatgacct gacaaccctg atcatcatgg
     6601 acgtacagaa acaagatggt ggactttata ccctgagttt agggaatgaa tttggatctg
     6661 actctgccac tgtgaatata catattcgat ccatttaaga gggcctgtgc gaattcaatc
     6721 gaattcctgc agcccggggg atccgtcgac ctgcagccaa gctaattccg ggcgaatttc
     6781 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     6841 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
     6901 gtcaggttgc tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc
     6961 atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7021 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7081 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7141 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7201 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7261 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7321 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7381 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7441 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7501 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7561 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7621 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7681 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7741 ctagtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7801 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7861 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7921 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7981 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8041 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8101 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8161 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8221 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8281 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8341 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8401 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8461 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8521 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8581 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8641 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8701 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8761 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8821 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8881 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8941 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9001 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9061 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9121 gaaaagtgcc acct
//
