LOCUS       Exported                8274 bp DNA     circular SYN 15-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8274)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8274)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..734
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1251..1280
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             complement(1464..1475)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     terminator      2309..2356
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2383..2570
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2594..2610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2618..2634)
                     /label=lac operator
                     /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(2642..2672)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2687..2708)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2996..3584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(3913..3960)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(4195..4986)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4987..5091)
                     /label=AmpR promoter
     rep_origin      complement(5118..6460)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        6719..7000
                     /label=TRP1 promoter
     CDS             7001..7672
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(7778..8233)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
       61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
      121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
      181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
      241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
      301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
      361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
      421 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
      481 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
      541 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
      601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
      721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
      781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
     1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
     1261 tgatctcaga ggaggacctg catatggcca tggaggccga attccctgtc accgagaccc
     1321 ctgggccagt ggcccctgcc ccagccactc catggcccct gtcatctttt gtcccttctc
     1381 aaaaaactta ccagggcaac tatggcttcc acctgggctt cctgcagtct gggacagcca
     1441 agtctgttat gtgcacgtac tctcctcccc tcaataagct attctgccag ctggcgaaga
     1501 cgtgccctgt gcagttgtgg gtcagcgcca cacctccagc tgggagccgt gtccgcgcca
     1561 tggccatcta caagaagtca cagcacatga cggaggtcgt gagacgctgc ccccaccatg
     1621 agcgctgctc cgatggtgat ggcctggctc ctccccagca tcttatccgg gtggaaggaa
     1681 atttgtatcc cgagtatctg gaagacaggc agacttttcg ccacagcgtg gtggtacctt
     1741 atgagccacc cgaggccggc tctgagtata ccaccatcca ctacaagtac atgtgtaata
     1801 gctcctgcat ggggggcatg aaccgccgac ctatccttac catcatcaca ctggaagact
     1861 ccagtgggaa ccttctggga cgggacagct ttgaggttcg tgtttgtgcc tgccctggga
     1921 gagaccgccg tacagaagaa gaaaatttcc gcaaaaagga agtcctttgc cctgaactgc
     1981 ccccagggag cgcaaagaga gcgctgccca cctgcacaag cgcctctccc ccgcaaaaga
     2041 aaaaaccact tgatggagag tatttcaccc tcaagatccg cgggcgtaaa cgcttcgaga
     2101 tgttccggga gctgaatgag gccttagagt taaaggatgc ccatgctaca gaggagtctg
     2161 gagacagcag ggctcactcc agctacctga agaccaagaa gggccagtct acttcccgcc
     2221 ataaaaaaac aatggtcaag aaagtggggc ctgactcaga ctgactgcct ctgcatcccg
     2281 gatccgtcga cctgcagcgg ccgcataact agcataaccc cttggggcct ctaaacgggt
     2341 cttgaggggt tttttgcgcg cttgcagcca agctaattcc gggcgaattt cttatgattt
     2401 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     2461 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
     2521 ctttctcagg tatagcatga ggtcgctctt attgaccaca cctctaccgg catgcaagct
     2581 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2641 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2701 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2761 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2821 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2881 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2941 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3001 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3061 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3121 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3181 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3241 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3301 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3361 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3421 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3481 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3541 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3601 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3661 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3721 tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg
     3781 cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg
     3841 cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa
     3901 ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg
     3961 tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag
     4021 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     4081 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg
     4141 atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa
     4201 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     4261 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     4321 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     4381 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgagtca cgacgagatc
     4441 ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     4501 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     4561 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     4621 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     4681 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     4741 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     4801 gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     4861 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     4921 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     4981 aatcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5041 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5101 ccgaaaagtg ccacctgaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg
     5161 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     5221 acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg
     5281 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     5341 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     5401 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     5461 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     5521 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     5581 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     5641 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     5701 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     5761 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     5821 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     5881 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     5941 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt
     6001 agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt
     6061 ttggttttca aaagcgctct gaagttccta tactttctag agaataggaa cttcggaata
     6121 ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca
     6181 tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg
     6241 agaagaacgg catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt
     6301 aggatgaaag gtagtctagt acctcctgtg atattatccc attccatgcg gggtatcgta
     6361 tgcttccttc agcactaccc tttagctgtt ctatatgctg ccactcctca attggattag
     6421 tctcatcctt caatgctatc atttcctttg atattggatc atactaagaa accattatta
     6481 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     6541 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     6601 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     6661 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catagatcaa
     6721 cgacattact atatatataa tataggaagc atttaataga acagcatcgt aatatatgtg
     6781 tactttgcag ttatgacgcc agatggcagt agtggaagat attctttatt gaaaaatagc
     6841 ttgtcacctt acgtacaatc ttgatccgga gcttttcttt ttttgccgat taagaattaa
     6901 ttcggtcgaa aaaagaaaag gagagggcca agagggaggg cattggtgac tattgagcac
     6961 gtgagtatac gtgattaagc acacaaaggc agcttggagt atgtctgtta ttaatttcac
     7021 aggtagttct ggtccattgg tgaaagtttg cggcttgcag agcacagagg ccgcagaatg
     7081 tgctctagat tccgatgctg acttgctggg tattatatgt gtgcccaata gaaagagaac
     7141 aattgacccg gttattgcaa ggaaaatttc aagtcttgta aaagcatata aaaatagttc
     7201 aggcactccg aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg atgttttggc
     7261 tctggtcaat gattacggca ttgatatcgt ccaactgcat ggagatgagt cgtggcaaga
     7321 ataccaagag ttcctcggtt tgccagttat taaaagactc gtatttccaa aagactgcaa
     7381 catactactc agtgcagctt cacagaaacc tcattcgttt attcccttgt ttgattcaga
     7441 agcaggtggg acaggtgaac ttttggattg gaactcgatt tctgactggg ttggaaggca
     7501 agagagcccc gaaagcttac attttatgtt agctggtgga ctgacgccag aaaatgttgg
     7561 tgatgcgctt agattaaatg gcgttattgg tgttgatgta agcggaggtg tggagacaaa
     7621 tggtgtaaaa gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga aataggttat
     7681 tactgagtag tatttattta agtattgttt gtgcacttgc cgatctatgc ggtgtgaaat
     7741 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     7801 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     7861 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     7921 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     7981 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     8041 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     8101 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     8161 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     8221 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgat
//
