LOCUS       sequence-11860-e        8506 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 8506)
  AUTHORS   Losev E, Reinke CA, Jellen J, Strongin DE, Bevis BJ, Glick BS
  TITLE     Golgi maturation visualized in living yeast.
  JOURNAL   Nature. 2006 Jun 22. 441(7096):1002-6.
  PUBMED    16699524
REFERENCE   2  (bases 1 to 8506)
  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: "Nature. 
            2006 Jun 22. 441(7096):1002-6."
COMMENT     Sequence Label: YCPlac33-sGFP-VRG4
FEATURES             Location/Qualifiers
     source          1..8506
                     /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"
     CDS             726..1439
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MGKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(756..784)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1379..1399
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3193..3210)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3193..3209)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3202..3224)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    3968..5130
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      4293..5130
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(5465..6268)
                     /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"
     promoter        complement(6269..6489)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6514..6532)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6600..6704
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6705..6773,6774..7565)
                     /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(6923..6942)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7736..8324
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8225..8244
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8478..8495
                     /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 ccaagcttgt
      241 aatggaccat ggcaatgtgc gccatatata tatgcggctc ccttatgagt agaagaaaat
      301 atatgactgg tggtacaaag tggcatggtg gtatatagat gtaggaacac catttgagtt
      361 tgccataatg attaaatttc cctttttata aagaattagc cgcccataac acgaaaaata
      421 tcacagttga cgaaagaaga cacgtcgcct aaacgcgtaa tgaatcgcaa tatacgaaaa
      481 gctaaacggg tttttcagaa tcctcttacc cagccttgga taatagtttt ggccgaaaaa
      541 aaaaaaaaga aagaaaaaaa ttgaaatttc gcgatccgaa caaacaatga acagaaaaac
      601 agaatatata taacgctgcc aagcctattt cttgttgccg attaattgtt gacatcctct
      661 taattaccaa aagagcctaa gaaaacaaac acactaacca cacagtatct ttcgcccgaa
      721 tgagcatggg taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
      781 atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat
      841 acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt ccttggccaa
      901 cacttgtcac tactttcact tatggtgttc agtgcttttc aagatacccg gatcatatga
      961 aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat
     1021 ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc
     1081 ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac
     1141 acaaattgga atacaactat aactcacaca atgtatacat catggcagac aaacaaaaga
     1201 atggaatcaa agctaacttc aaaattagac acaacattga agatggaagc gttcaactag
     1261 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
     1321 attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg
     1381 tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaat
     1441 catgctcaga attgaaaaca ggtcatgcag gccataaccc ttgggcttca gttgccaatt
     1501 ccggtccgat ctctatttta tcctactgtg gttcctctat tttaatgacg gtgactaaca
     1561 agttcgtcgt caatttgaag gatttcaaca tgaactttgt catgcttttc gtgcaatctt
     1621 tggtttgtac tataaccttg attatcctac gtatactggg ctatgcgaag ttccgttcat
     1681 taaacaaaac agacgccaag aactggttcc ctatttcctt tttactggtc ttgatgatct
     1741 acacctcttc gaaggcttta caatacttgg ctgttccaat ttacaccatt ttcaagaatt
     1801 tgactattat cttgattgct tatggtgagg ttctcttttt tggtggctct gtcacctcca
     1861 tggaattgtc atcatttttg ttgatggtcc tttcttctgt cgttgcaact tggggtgacc
     1921 agcaagctgt ggctgccaag gctgcttcat tggctgaagg agcagccggt gctgttgcct
     1981 cctttaaccc aggttatttc tggatgttca ccaactgtat cacttctgca ttattcgttc
     2041 ttataatgag aaagagaatt aagttaacta acttcaagga tttcgacact atgttttaca
     2101 acaatgtttt ggctctacct attctattgc tgttttcttt ctgtgtggaa gattggtctt
     2161 cagttaattt gaccaataac ttttctaacg attcgctaac tgctatgatc atcagtggtg
     2221 ttgcatccgt cggtatttct tactgttccg gttggtgtgt tcgtgttact tcgtctacta
     2281 catattcgat ggtaggggct ttgaacaagc tgccaattgc cttgtctggt ttgattttct
     2341 ttgatgctcc aagaaacttc ttatctattc tctccatttt tattggtttc ctatcaggta
     2401 ttatttatgc tgttgccaaa caaaagaagc aacaagccca acctttacgt aaatgagaac
     2461 ttacgggggg tgcaatttat tttttttttt tggtttattt tattttatag aggcatctaa
     2521 tgcaagtaga tttatataca attatactta aaattgatat accttagaac aggtcggaca
     2581 caacgtgtgg taactccaat atatcactaa tgacaatagc gccttctggt gtttgcccca
     2641 aaatttcatt tacttcattc tcaactaaat gaatacatgt tttcatgccc aatttgatac
     2701 ctgtctcgat gtttttccca ctgtcatcta taaaataagc attctcgtat cttgccaacc
     2761 cgctttcctt catggctttt tcaaatgctt taacatgtgg tttgcagacc agcgtatctg
     2821 tcctagagta gtcacaatat gtcaatccat caaataaatc cgctataccc aacaatctca
     2881 ggcaacgtat tgcgtggttt ttataagcgt tggtgaaaag ccacaattta tcaatctttc
     2941 ccgattgtct caatctcaac agcatattcc ttaatggtat atcaggtttc aagatatctt
     3001 gtagtggtaa ggaatcatcc acaagccggt tatattccag tgcgttcact ttgtggaaca
     3061 ttacaagtcc ccttatggca agtccatact ctttgtagta agagttattc aaaacgtgag
     3121 catcttcagg tgatagtttc aagtgtgttt gaaagaacct cagtatagat tgttgcataa
     3181 gatcgtgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3241 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3301 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     3361 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatatcgct gggccattct
     3421 catgaagaat atcttgaatt tattgtcata ttactagttg gtgtggaagt ccatatatcg
     3481 gtgatcaata tagtggttga catgctggct agtcaacatt gagccttttg atcatgcaaa
     3541 tatattacgg tattttacaa tcaaatatca aacttaacta ttgactttat aacttattta
     3601 ggtggtaaca ttcttataaa aaagaaaaaa attactgcaa aacagtacta gcttttaact
     3661 tgtatcctag gttatctatg ctgtctcacc atagagaata ttacctattt cagaatgtat
     3721 gtccatgatt cgccgggtaa atacatataa tacacaaatc tggcttaata aagtctataa
     3781 tatatctcat aaagaagtgc taaattggct agtgctatat atttttaaga aaatttcttt
     3841 tgactaagtc catatcgact ttgtaaaagt tcactttagc atacatatat tacacgagcc
     3901 agaaattgta acttttgcct aaaatcacaa attgcaaaat ttaattgctt gcaaaaggtc
     3961 acatgcttat aatcaacttt tttaaaaatt taaaatactt ttttattttt tatttttaaa
     4021 cataaatgaa ataatttatt tattgtttat gattaccgaa acataaaacc tgctcaagaa
     4081 aaagaaactg ttttgtcctt ggaaaaaaag cactacctag gagcggccaa aatgccgagg
     4141 ctttcatagc ttaaactctt tacagaaaat aggcattata gatcagttcg agttttctta
     4201 ttcttccttc cggttttatc gtcacagttt tacagtaaat aagtatcacc tcttagagtt
     4261 cgatgataag ctgtcaaaca tgagaattaa ttccacatgt taaaatagtg aaggagcatg
     4321 ttcggcacac agtggaccga acgtggggta agtgcactag ggtccggtta aacggatctc
     4381 gcattgatga ggcaacgcta attatcaaca tatagattgt tatctatctg catgaacacg
     4441 aaatctttac ttgacgactt gaggctgatg gtgtttatgc aaagaaacca ctgtgtttaa
     4501 tatgtgtcac tgtttgatat tactgtcagc gtagaagata atagtaaaag cggttaataa
     4561 gtgtatttga gataagtgtg ataaagtttt tacagcgaaa agacgataaa tacaagaaaa
     4621 tgattacgag gatacggaga gaggtatgta catgtgtatt tatatactaa gctgccggcg
     4681 gttgtttgca agaccgagaa aaggctagca agaatcgggt cattgtagcg tatgcgcctg
     4741 tgaacattct cttcaacaag tttgattcca ttgcggtgaa atggtaaaag tcaaccccct
     4801 gcgatgtata ttttcctgta caatcaatca aaaagccaaa tgatttagca ttatctttac
     4861 atcttgttat tttacagatt ttatgtttag atcttttatg cttgcttttc aaaaggcttg
     4921 caggcaagtg cacaaacaat acttaaataa atactactca gtaataacct atttcttagc
     4981 atttttgacg aaatttgcta ttttgttaga gtcttttaca ccatttgtct ccacacctcc
     5041 gcttacatca acaccaataa cgccatttaa tctaagcgca tcaccaacat tttctggcgt
     5101 cagtccacca gctaacataa aatgtaagct ctcggggctc tcttgccttc caacccagtc
     5161 agaaatcgag ttccaatcca aaagttcacc tgtcccacct gcttctgaat caaacaaggg
     5221 aataaacgaa tgaggtttct gtgaagctgc actgagtagt atgttgcagt cttttggaaa
     5281 tacgagtctt ttaataactg gcaaaccgag gaactcttgg tattcttgcc acgactcatc
     5341 tccatgcagt tggacgatcg atgataagct gtcaaacatg agaattgggt aataactgat
     5401 ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag
     5461 ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac
     5521 gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa
     5581 tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc
     5641 ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc
     5701 cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt
     5761 caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta
     5821 acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa
     5881 caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt
     5941 atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt
     6001 cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca
     6061 tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg
     6121 cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg
     6181 tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag
     6241 cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcat gtttttgttc
     6301 tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta
     6361 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg
     6421 aatcaaaaaa atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat
     6481 tgaattgaaa agctaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     6541 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     6601 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6661 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6721 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6781 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6841 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6901 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6961 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7021 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7081 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7141 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7201 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7261 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7321 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7381 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7441 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7501 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7561 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7621 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7681 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7741 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7801 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7861 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     7921 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7981 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8041 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8101 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8161 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8221 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8281 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8341 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8401 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     8461 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
