LOCUS       Exported                7209 bp DNA     circular SYN 21-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7209)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7209)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7209)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7209)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2510..2518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             152..811
                     /codon_start=1
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        2580..2861
                     /label=TRP1 promoter
     CDS             2862..3536
                     /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"
     CDS             2862..3533
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3618..3636)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3657..3673)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3681..3697)
                     /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(3705..3735)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3750..3771)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4059..4647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4818..5612)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5613..5717)
                     /label=AmpR promoter
     misc_feature    complement(5750..6253)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     rep_origin      complement(6570..7025)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     7166..7182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7189..7207
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaattcccgg ggagctcacg cgttcgcgaa tcgatccgcg gtctagaaat tcctggcatt
       61 atcacataat gaattataca ttatataaag taatgtgatt tcttcgaaga atatactaaa
      121 aaatgagcag gcaagataaa cgaaggcaaa gatgacagag cagaaagccc tagtaaagcg
      181 tattacaaat gaaaccaaga ttcagattgc gatctcttta aagggtggtc ccctagcgat
      241 agagcactcg atcttcccag aaaaagaggc agaagcagta gcagaacagg ccacacaatc
      301 gcaagtgatt aacgtccaca caggtatagg gtttctggac catatgatac atgctctggc
      361 caagcattcc ggctggtcgc taatcgttga gtgcattggt gacttacaca tagacgacca
      421 tcacaccact gaagactgcg ggattgctct cggtcaagct tttaaagagg ccctaggggc
      481 cgtgcgtgga gtaaaaaggt ttggatcagg atttgcgcct ttggatgagg cactttccag
      541 agcggtggta gatctttcga acaggccgta cgcagttgtc gaacttggtt tgcaaaggga
      601 gaaagtagga gatctctctt gcgagatgat cccgcatttt cttgaaagct ttgcagaggc
      661 tagcagaatt accctccacg ttgattgtct gcgaggcaag aatgatcatc accgtagtga
      721 gagtgcgttc aaggctcttg cggttgccat aagagaagcc acctcgccca atggtaccaa
      781 cgatgttccc tccaccaaag gtgttcttat gtagtgacac cgattattta aagctgcagc
      841 atacgatata tatacatgtg tatatatgta tacctatgaa tgtcagtaag tatgtatacg
      901 aacagtatga tactgaagat gacaaggtaa tgcatcattc tatacgtgtc attctgaacg
      961 aggcgcgctt tccttttttc tttttgcttt ttcttttttt ttctcttgaa ctcgagaaaa
     1021 aaaatataaa agagatggag gaacgggaaa aagttagttg tggtgatagg tggcaagtgg
     1081 tattccgtaa gaacaacaag aaaagcattt catattatgg ctgaactgag cgaacaagtg
     1141 caaaatttaa gcatcaacga caacaacgag aatggttatg ttcctcctca cttaagagga
     1201 aaaccaagaa gtgccagaaa taacagtagc aactacaata acaacaacgg cggctacaac
     1261 ggtggccgtg gcggtggcag cttctttagc aacaaccgtc gtggtggtta cggcaacggt
     1321 ggtttcttcg gtggaaacaa cggtggcagc agatctaacg gccgttctgg tggtagatgg
     1381 atcgatggca aacatgtccc agctccaaga aacgaaaagg ccgagatcgc catatttggt
     1441 gtccccgagg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     1501 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     1561 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     1621 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     1681 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgac cgatgccctt
     1741 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     1801 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     1861 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     1921 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     1981 cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct
     2041 ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg
     2101 catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg
     2161 acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcattg gaccgctgat
     2221 cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg
     2281 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     2341 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga
     2401 gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc
     2461 atcgcgtccg ccatctccag cagccgcacg cggcgcatcg gggggggggg ggggggggtt
     2521 tcaattcatc attttttttt tattcttttt tttgatttcg gtttctttga aattttttta
     2581 acgacattac tatatatata atataggaag catttaatag aacagcatcg taatatatgt
     2641 gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat tgaaaaatag
     2701 cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga ttaagaatta
     2761 attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga ctattgagca
     2821 cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt attaatttca
     2881 caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag gccgcagaat
     2941 gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat agaaagagaa
     3001 caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt
     3061 caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg
     3121 ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcaag
     3181 aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca
     3241 acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag
     3301 aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc
     3361 aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca gaaaatgttg
     3421 gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa
     3481 atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta
     3541 ttactgagta gtatttattt aagtattgtt tgtgcacttg ccctagagtc gacctgcagg
     3601 catgcaagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca
     3661 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3721 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3781 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4801 taaacttggt ctgacagtca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     4861 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     4921 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     4981 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     5041 gcatcgccat gagtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     5101 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     5161 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     5221 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     5281 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     5341 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcagggaac gcccgtcgtg
     5401 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     5461 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     5521 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     5581 gaacctgcgt gcaatccatc ttgttcaatc atactcttcc tttttcaata ttattgaagc
     5641 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5701 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcggacgg atcgcttgcc
     5761 tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg
     5821 tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag
     5881 ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact
     5941 ttacatatat atttattaga caagaaaagc agattaaata gatatacatt cgattaacga
     6001 taagtaaaat gtaaaatcac aggattttcg tgtgtggtct tctacacaga caagatgaaa
     6061 caattcggca ttaatacctg agagcaggaa gagcaagata aaaggtagta tttgttggcg
     6121 atccccctag agtcttttac atcttcggaa aacaaaaact attttttctt taatttcttt
     6181 ttttactttc tatttttaat ttatatattt atattaaaaa atttaaatta taattatttt
     6241 tatagcacgt gatgaaaagg accgacgtct aagaaaccat tattatcatg acattaacct
     6301 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6361 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6421 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6481 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     6541 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     6601 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6661 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6721 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6781 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6841 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6901 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6961 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     7021 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     7081 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     7141 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
     7201 ctatagggc
//
