LOCUS       40924_21225        5524 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5524)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5524)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..406
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             434..874
                     /codon_start=1
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     terminator      921..1108
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1164..1835)
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(1836..1937)
                     /label=TRP1 promoter
     primer_bind     complement(1952..1968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1976..1992)
                     /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(2000..2030)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2045..2066)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2354..2942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3116..3973)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3974..4078)
                     /label=AmpR promoter
     rep_origin      4360..5524
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga
      121 cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatcttcgaa cacacgaaac
      181 tttttccttc cttcattcac gcacactact ctctaatgag caacggtata cggccttcct
      241 tccagttact tgaatttgaa ataaaaaaag tttgccgctt tgctatcaag tataaataga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaagca
      421 agcctcctga aagatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
      481 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
      541 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
      601 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
      661 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
      721 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
      781 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
      841 tagtaacaaa ggtcaaagac agttgactgt atcgccggaa ttcccgggga tccgtcgacc
      901 tgcagccaag ctaattccgg gcgaatttct tatgatttat gatttttatt attaaataag
      961 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     1021 tcttgttctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
     1081 tcgctcttat tgaccacacc tctaccggca tgccggcaag tgcacaaaca atacttaaat
     1141 aaatactact cagtaataac ctatttctta gcatttttga cgaaatttgc tattttgtta
     1201 gagtctttta caccatttgt ctccacacct ccgcttacat caacaccaat aacgccattt
     1261 aatctaagcg catcaccaac attttctggc gtcagtccac cagctaacat aaaatgtaag
     1321 ctttcggggc tctcttgcct tccaacccag tcagaaatcg agttccaatc caaaagttca
     1381 cctgtcccac ctgcttctga atcaaacaag ggaataaacg aatgaggttt ctgtgaagct
     1441 gcactgagta gtatgttgca gtcttttgga aatacgagtc ttttaataac tggcaaaccg
     1501 aggaactctt ggtattcttg ccacgactca tctccatgca gttggacgat atcaatgccg
     1561 taatcattga ccagagccaa aacatcctcc ttaggttgat tacgaaacac gccaaccaag
     1621 tatttcggag tgcctgaact atttttatat gcttttacaa gacttgaaat tttccttgca
     1681 ataaccgggt caattgttct ctttctattg ggcacacata taatacccag caagtcagca
     1741 tcggaatcta gagcacattc tgcggcctct gtgctctgca agccgcaaac tttcaccaat
     1801 ggaccagaac tacctgtgaa attaataaca gacatactcc aagctgcctt tgtgtgctta
     1861 atcacgtata ctcacgtgct caatagtcac caatgccctc cctcttggcc ctctcctttt
     1921 cttttttcga ccgaattaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1981 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2041 cctaatgagt gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2101 gaaacctgtc gtgccagctg gattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2161 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2221 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2281 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2341 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2401 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2461 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2521 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2581 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2641 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2701 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2761 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     2821 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2881 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3181 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3241 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3301 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3361 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3421 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3481 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3541 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3601 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3661 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3721 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3781 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3841 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3901 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3961 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4021 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4081 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4141 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     4201 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4261 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     4321 atgcggcatc agagcagatt gtactgagag tgcaccataa cgcatttaag cataaacacg
     4381 cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac
     4441 gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg
     4501 aaacgctttg aagttcctat tccgaagttc ctattctcta gctagaaagt ataggaactt
     4561 cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc
     4621 accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac attgctcaaa
     4681 agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt
     4741 cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt
     4801 actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg
     4861 aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa
     4921 ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat
     4981 ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct
     5041 tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata
     5101 gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag
     5161 actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa
     5221 tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa
     5281 aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
     5341 aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt
     5401 ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg
     5461 tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc acagatgctt
     5521 cgtt
//
