LOCUS       Exported                6552 bp DNA     circular SYN 19-SEP-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6552)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6552)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6552)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          8..14
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(5..259)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(322..1035)
                     /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(1067..1085)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1117..1558)
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     rep_origin      1580..2093
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2104..2981
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3582..3797
                     /label=URA3 promoter
     CDS             3798..4601
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             4697..5557
                     /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"
     rep_origin      5728..6316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggccgcagct tgcaaattaa agccttcgag cgtcccaaaa ccttctcaag caaggttttc
       61 agtataatgt tacatgcgta cacgcgtctg tacagaaaaa aaagaaaaat ttgaaatata
      121 aataacgttc ttaatactaa cataactata aaaaaataaa tagggaccta gacttcaggt
      181 tgtctaactc cttccttttc ggttagagcg gatgtggggg gagggcgtga atgtaagcgt
      241 gacataacta attacatgat gcggccctct agatgcatgc tcgagcggcc gccagtgtga
      301 tggatatctg cagaattctt atttgtatag ttcatccatg ccatgtgtaa tcccagcagc
      361 tgttacaaac tcaagaagga ccatgtggtc tctcttttcg ttgggatctt tcgaaagggc
      421 agattgtgtg gacaggtaat ggttgtctgg taaaaggaca gggccatcgc caattggagt
      481 attttgttga taatggtctg ctagttgaac gcttccatct tcaatgttgt gtctaatttt
      541 gaagttaact ttgattccat tcttttgttt gtctgccatg atgtatacgt tgtgggagtt
      601 gtagttgtat tccaacttgt ggccgaggat gtttccgtcc tccttgaaat cgattccctt
      661 aagctcgatt ctgttgacga gggtgtctcc ctcaaacttg acttcagcac gtgtcttgta
      721 gttcccgtcg tccttgaaga agatggtcct ctcctgcacg tatccctcag gcatggcgct
      781 cttgaagaag tcgtgccgct tcatatgatc tgggtatctt gaaaagcatt gaacaccata
      841 agtaagagta gtgacaagtg ttggccaagg aacaggaagc tttccagtag tgcaaataaa
      901 tttaagggta agttttccgt atgttgcatc accttcaccc tctccactga cagaaaattt
      961 gtgcccatta acatcaccat ctaattcaac aagaattggg acaactccag tgaaaagttc
     1021 ttctccctta cccatggatc cgagctcggt accaagctta atattcccta tagtgagtcg
     1081 tattacagct gctagtagtc cgatccgggg ttttttctcc ttgacgttaa agtatagagg
     1141 tatattaaca attttttgtt gatactttta ttacatttga ataagaagta atacaaaccg
     1201 aaaatgttga aagtattagt taaagtggtt atgcagtttt tgcatttata tatctgttaa
     1261 tagatcaaaa atcatcgctt cgctgattaa ttaccccaga aataaggcta aaaaactaat
     1321 cgcattatca tcctatggtt gttaatttga ttcgttcatt tgaaggtttg tggggccagg
     1381 ttactgccaa tttttcctct tcataaccat aaaagctagt attgtagaat ctttattgtt
     1441 cggagcagtg cggcgcgagg cacatctgcg tttcaggaac gcgaccggtg aagacgagga
     1501 cgcacggagg agagtcttcc ttcggagggc tgtcacccgc tcggcggctt ctaatccgtg
     1561 gactagtgga tcgatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1621 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1681 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1741 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1801 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1861 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1921 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1981 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaaata
     2041 tttgcttata caatcttcct gtttttgggg cttttctgat tatcaaccgg ggtggagctt
     2101 cccattgcga ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc
     2161 tgtgctatat ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa
     2221 ctcgacctct acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta
     2281 gtaagaacta ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg
     2341 tagtatatag agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat
     2401 caacgctatc actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc
     2461 ggggatgcct ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg
     2521 gaagtggagt caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta
     2581 accttaacgg acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag
     2641 gagaaaaaaa gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt
     2701 gtagaacaaa aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt
     2761 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca
     2821 tttttgtttt acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg
     2881 catttctgtt ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg
     2941 ttgcattttt gttctacaaa atgaagcaca gatgcttcgt taacaaagat atgctattga
     3001 agtgcaagat ggaaacgcag aaaatgaacc ggggatgcga cgtgcaagat tacctatgca
     3061 atagatgcaa tagtttctcc aggaaccgaa atacatacat tgtcttccgt aaagcgctag
     3121 actatatatt attatacagg ttcaaatata ctatctgttt cagggaaaac tcccaggttc
     3181 ggatgttcaa aattcaatga tgggtaacaa gtacgatcgt aaatctgtaa aacagtttgt
     3241 cggatattag gctgtatctc ctcaaagcgt attcgaatat cattgagaag ctgcagcgtc
     3301 acatcggata ataatgatgg cagccattgt agaagtgcct tttgcatttc tagtctcttt
     3361 ctcggtctag ctagttttac tacatcgcga agatagaatc ttagatcaca ctgcctttgc
     3421 tgagctggat caatagagta acaaaagagt ggtaaggcct cgttaaagga caaggacctg
     3481 agcggaagtg tatcgtacag tagacggagt atactaggta tagtctatag tccgtggaat
     3541 taattctcat gtttgacagc ttatcatcga taagctagct tttcaattca tcattttttt
     3601 tttattcttt tttttgattt cggtttcctt gaaatttttt tgattcggta atctccgaac
     3661 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     3721 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacctgca
     3781 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     3841 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     3901 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     3961 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     4021 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaagac agaaaatttg
     4081 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     4141 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     4201 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     4261 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     4321 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     4381 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagac gcattgggtc
     4441 aacagtatag aaccgtggat gatgtggtct ctacaggatc tgacattatt attgttggaa
     4501 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     4561 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     4621 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattaccca
     4681 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4741 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4801 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4861 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4921 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4981 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5041 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5101 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5161 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5221 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5281 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5341 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5401 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5461 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5521 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5581 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5641 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5701 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5761 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5821 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5881 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5941 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6001 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6061 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6121 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6181 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6241 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6301 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6361 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6421 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6481 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6541 tcattaatgc ag
//
