LOCUS       sequence-9987-ex        5003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5003)
  AUTHORS   Rossanese OW, Reinke CA, Bevis BJ, Hammond AT, Sears IB, O'Connor J,
            Glick BS
  TITLE     A role for actin, Cdc1p, and Myo2p in the inheritance of late Golgi 
            elements in Saccharomyces cerevisiae.
  JOURNAL   J Cell Biol. 2001 Apr 2. 153(1):47-62.
  PUBMED    11285273
REFERENCE   2  (bases 1 to 5003)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol. 2001 Apr 2. 153(1):47-62."
COMMENT     Sequence Label: YIPlac204TKC-GFP-HDEL
FEATURES             Location/Qualifiers
     source          1..5003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(239..486)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     452..470
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(join(512..1222,1223..1225,1226..1228))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(512..1222,1223..1225,1226..1228))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(554..575)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     903..922
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1162..1183
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1225..1234
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1385..1904)
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     primer_bind     complement(1973..1992)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2092..2114
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2233..2907)
                     /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"
     promoter        complement(2908..3009)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3011..3029)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3097..3201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3202..3270,3271..4062)
                     /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"
     primer_bind     complement(3420..3439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4233..4821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4722..4741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4975..4992
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac
      301 atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta
      361 atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt
      421 ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac ataactaatt
      481 acatgatgac tctagattac aattcgtcgt gcttgtacag ctcgtccatg ccgagagtga
      541 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
      601 tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
      661 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
      721 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
      781 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
      841 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
      901 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
      961 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     1021 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     1081 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     1141 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     1201 tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tggatcccca tcatcggcac
     1261 ctctaactaa aacattggag gagtggaaag aattctgtaa aggtaatatt accacgaaaa
     1321 gggcgtacag gaccacggag agtggtacca gcagcttgcc agcgcttagt ctgttgaaaa
     1381 acatttttag tttatgtatg tgttttttgt agttatagat ttaagcaaga aaagaataca
     1441 aacaaaaaat tgaaaaagat tgatttagaa ttaaaaagaa aaatatttac gtaagaaggg
     1501 aaaatagtaa atgttgcaag ttcactaaac tcctaaatta tgctgccctt tatattccct
     1561 gttacagcag ccgagccaaa ggtatatagg ctcctttgca ttagcatgcg taacaaacca
     1621 cctgtcagtt tcaaccgagg tggtatccga gagaattgtg tgattgcttt aattaatttc
     1681 ggagaatctc acatgccact gaagattaaa aactggatgc cagaaaaggg gtgtccaggt
     1741 gtaacatcaa tagaggaagc tgaaaagtct tagaacgggt aatcttccac caacctgatg
     1801 ggttcctaga tataatctcg aagggaataa gtagggtgat accgcagaag tgtctgaatg
     1861 tattaaggtc ctcacagttt aaatcccgct cacactaacg taggcgcctg atgcggtatt
     1921 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     1981 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     2041 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2101 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcgatcttt
     2161 tatgcttgct tttcaaaagg cttgcaggca agtgcacaaa caatacttaa ataaatacta
     2221 ctcagtaata acctatttct tagcattttt gacgaaattt gctattttgt tagagtcttt
     2281 tacaccattt gtctccacac ctccgcttac atcaacacca ataacgccat ttaatctaag
     2341 cgcatcacca acattttctg gcgtcagtcc accagctaac ataaaatgta agctctcggg
     2401 gctctcttgc cttccaaccc agtcagaaat cgagttccaa tccaaaagtt cacctgtccc
     2461 acctgcttct gaatcaaaca agggaataaa cgaatgaggt ttctgtgaag ctgcactgag
     2521 tagtatgttg cagtcttttg gaaatacgag tcttttaata actggcaaac cgaggaactc
     2581 ttggtattct tgccacgact catctccatg cagttggacg atatcaatgc cgtaatcatt
     2641 gaccagagcc aaaacatcct ccttaggttg attacgaaac acgccaacca agtatttcgg
     2701 agtgcctgaa ctatttttat atgcttttac aagacttgaa attttccttg caataaccgg
     2761 gtcaattgtt ctctttctat tgggcacaca tataataccc agcaagtcag catcggaatc
     2821 tagtgcacat tctgcggcct ctgtgctctg caagccgcaa actttcacca atggaccaga
     2881 actacctgtg aaattaataa cagacatact ccaagctgcc tttgtgtgct taatcacgta
     2941 tactcacgtg ctcaatagtc accaatgccc tccctcttgg ccctctcctt ttcttttttc
     3001 gaccgaattg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3061 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3121 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3181 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3241 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3301 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3361 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3421 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3481 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3541 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3601 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3661 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3721 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3781 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3841 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3901 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3961 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4021 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4081 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4141 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4201 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4261 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4321 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     4381 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4441 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4501 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4561 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4621 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4681 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4741 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4801 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4861 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4921 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4981 tcagtgagcg aggaagcgga aga
//
