LOCUS       40924_48028        6548 bp DNA     circular SYN 11-SEP-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6548)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6548)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(4..251)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(314..1027)
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="MGKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     promoter        complement(1059..1077)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1109..1551)
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     rep_origin      1570..2083
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2094..2974
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3573..3793
                     /label=URA3 promoter
     CDS             3794..4594
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     misc_feature    4877..5255
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     rep_origin      5724..6312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccgcaaatt aaagccttcg agcgtcccaa aaccttctca agcaaggttt tcagtataat
       61 gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa atttgaaata taaataacgt
      121 tcttaatact aacataacta taaaaaaata aatagggacc tagacttcag gttgtctaac
      181 tccttccttt tcggttagag cggatgtggg gggagggcgt gaatgtaagc gtgacataac
      241 taattacatg atgcggccct ctagatgcat gctcgagcgg ccgccagtgt gatggatatc
      301 tgcagaattc ttatttgtat agttcatcca tgccatgtgt aatcccagca gctgttacaa
      361 actcaagaag gaccatgtgg tctctctttt cgttgggatc tttcgaaagg gcagattgtg
      421 tggacaggta atggttgtct ggtaaaagga cagggccatc gccaattgga gtattttgtt
      481 gataatggtc tgctagttga acgcttccat cttcaatgtt gtgtctaatt ttgaagttaa
      541 ctttgattcc attcttttgt ttgtctgcca tgatgtatac gttgtgggag ttgtagttgt
      601 attccaactt gtggccgagg atgtttccgt cctccttgaa atcgattccc ttaagctcga
      661 ttctgttgac gagggtgtct ccctcaaact tgacttcagc acgtgtcttg tagttcccgt
      721 cgtccttgaa gaagatggtc ctctcctgca cgtatccctc aggcatggcg ctcttgaaga
      781 agtcgtgccg cttcatatga tctgggtatc ttgaaaagca ttgaacacca taagtaagag
      841 tagtgacaag tgttggccaa ggaacaggaa gctttccagt agtgcaaata aatttaaggg
      901 taagttttcc gtatgttgca tcaccttcac cctctccact gacagaaaat ttgtgcccat
      961 taacatcacc atctaattca acaagaattg ggacaactcc agtgaaaagt tcttctccct
     1021 tacccatgga tccgagctcg gtaccaagct taatattccc tatagtgagt cgtattacag
     1081 ctgctagtag tccgatccgg ggttttttct ccttgacgtt aaagtataga ggtatattaa
     1141 caattttttg ttgatacttt tattacattt gaataagaag taatacaaac cgaaaatgtt
     1201 gaaagtatta gttaaagtgg ttaatgcagt ttttgcattt atatatctgt taatagatca
     1261 aaaatcatcg cttcgctgat taattacccc agaaataagg ctaaaaaact aatcgcatta
     1321 tcatcctatg gttgttaatt tgattcgttc atttgaaggt ttgtggggcc aggttactgc
     1381 caatttttcc tcttcataac cataaaagct agtattgtag aatctttatt gttcggagca
     1441 gtgcggcgcg aggcacatct gcgtttcagg aacgcgaccg gtgaggacga ggacgcacgg
     1501 aggagagtct tccttcggag ggctgtcacc cgctcggcgg cttctaatcc gtactagtgg
     1561 atcatcccca cgcgccctgt agcgccccat taagcgcggc gggtgtggtg gttacgccca
     1621 gcgtgacccc tacacttccc accgccctag cccccgctcc tttcgctttc ttcccttcct
     1681 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tgggggcatc cgtttaccct
     1741 tccgatttac tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1801 gtagtgggcc atcgccctga tagacccttt ttcgcccttt gacgttggag tccacgttct
     1861 ttaatagtgg actcttgttg gaaactggaa caacactcaa ccctatctcg gtctattctt
     1921 ttgatttata agggattttg ccgatttcgg gctattcgtt aaaaaatgag ctgatttaac
     1981 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaata tttgcttata
     2041 caatcttcct gtttttgggg cttttctgat tatcaaccgg ggtggagctt cccattgcga
     2101 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
     2161 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
     2221 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
     2281 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
     2341 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
     2401 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
     2461 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
     2521 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
     2581 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
     2641 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
     2701 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
     2761 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgtcgcgtt gcatttttgt
     2821 tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct
     2881 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt
     2941 tttgttctac aaaatgaagc acagatgctt cgttaacaaa gatatgctat tgaagtgcaa
     3001 gatggaaacg cagaaaatga accggggatg cgacgtgcaa gattacctat gcaatagatg
     3061 caatagtttc tccaggaacc gaaatacata cattgtcttc cgtaaagcgc tagactatat
     3121 attattatac aggttcaaat atactatctg tttcagggaa aactcccagg ttcggatgtt
     3181 caaaattcaa tgatgggtaa caagtacgat cgtaaatctg taaaacagtt tgtcggatat
     3241 taggctgtat ctcctcaaag cgtattcgaa tatcattgag aagctgcagc gtcacatcgg
     3301 ataataatga tggcagccat tgtagaagtg ccttttgcat ttctagtctc tttctcggtc
     3361 tagctagttt tactacatcg cgaagataga atcttagatc acactgcctt tgctgagctg
     3421 gatcaataga gtaacaaaag agtggtaagg cctcgttaaa ggacaaggac ctgagcggaa
     3481 gtgtatcgta cagtagacgg agtatctagt atagtctata gtccgtggaa ttaattctca
     3541 tctttgacag cttatcatcg ataagctagc ttttcaattc aattcatcat ttttttttta
     3601 ttcttttttt tgatttcggt ttctttgaaa tttttttgat tcggtaatct ccgaacagaa
     3661 ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga
     3721 agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa
     3781 acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc
     3841 tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt
     3901 ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg
     3961 tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc
     4021 gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa aatttgctga
     4081 cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc
     4141 agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc
     4201 ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa
     4261 gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa
     4321 agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga
     4381 ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat tgggtcaaca
     4441 gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg ttggaagagg
     4501 actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg
     4561 ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc
     4621 atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat tacccattga
     4681 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4741 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4801 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4861 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtgat
     4921 acactattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4981 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5041 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5101 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5161 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagagt
     5221 gacaccacga tgcctgtagc aatgccaaca acgttgcgca aactattaac tggcgaacta
     5281 cttactctag cttcccggca acaattaata gactgaatgg aggcggataa agttgcagga
     5341 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5401 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcg ctcccgtatc
     5461 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5521 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5581 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5641 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5701 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5761 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5821 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     5881 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5941 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6001 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6061 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6121 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6181 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6241 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6301 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctgggctt ttgctggcct
     6361 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6421 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6481 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6541 taatgcag
//
