LOCUS       sequence-11862-e        8290 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 8290)
  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 8290)
  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: pSSec7-mGFPx3
FEATURES             Location/Qualifiers
     source          1..8290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      988..997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(994..996,997..999,1000..1710)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      1720..1729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1726..1728,1729..1731,1732..2442)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2452..2461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2458..2460,2461..2463,2464..3177)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4907..4923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4907..4923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4920..4942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4931..4947
                     /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)."
     promoter        complement(join(4955..4961,4962..4979,4980..4985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5000..5021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5138..5155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5309..5897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5389..5408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6068..6859,6860..6928))
                     /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     6691..6710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6929..7033)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(7200..8003)
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(8004..8220)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     8263..8281
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ctattgttgt caaatgtgtt
      421 ttacagcttt tgatgataga gctgctcaac gaattattcg agaacgaaga ttttgcccac
      481 tgtatccctt acaaggaagc cattagaatt acaagattgt tggagaaatc gtacgaattt
      541 tctcgtgact ttaatgaaga ttatgggtta agaacaagac tagtagaggc tcgtgtagtt
      601 gataaaatac caaacctgtt gaaacaagaa acgagtgctg ctgcagttct ccttgacatt
      661 atgtttcagt tgtatctgaa cgatgatgag aagaaggctg atttaataac tcgtctgatc
      721 accatttgca tccaggtcgt agagggttat gtttctttgg atgatagaac catggaacgt
      781 agtatcaatg cctggcgttc agtgatagtt gaaatattac aaggttacta cgaatttgat
      841 gatgaagact ttagactata ctgtcctgct atgtatgctc tggtgattca aatattagat
      901 aagagcgttc caacggagtt gagacatgct ataaaacaat ttctaagcag agttggtgaa
      961 ttataccttt ctacggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc
     1021 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1081 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1141 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1201 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1261 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1321 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1381 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1441 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1501 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1561 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc
     1621 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1681 atcactctcg gcatggacga gctgtacaag ggtccggtcg ccaccatggt gagcaagggc
     1741 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1801 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1861 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1921 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     1981 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2041 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2101 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2161 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     2221 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2281 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     2341 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2401 accgccgccg ggatcactct cggcatggac gagctgtaca agggtccggt cgccaccatg
     2461 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2521 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2581 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2641 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2701 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2761 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2821 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2881 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2941 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     3001 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3061 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3121 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     3181 ggccgggctt tgttgaatgg tccagcagat taaaatatgc ttagttgtag aacacagaaa
     3241 aaatgagaag gttgatatac ttttactcat attaatcctg atttagtggc ggtgatcctt
     3301 tttagattcg tttaattcat aaacaaggta catcatatag caataacaaa tatgtaaact
     3361 atgtagaaag acttatcaag tttagccatt ttggtcctca ttcatgggca ttttatacta
     3421 acttcatttt tgccgaaata agaaaggatg taatcttaac ctaaaatatg taagcaatta
     3481 agggcgcacg tagtcacttg cgaacaaata ggaaaagaaa gaaacaagca cggatactga
     3541 atgactgagt gtattactac acgatttatc tcaacgaatt taagtgtgga ggacgatcgg
     3601 ctgcacgtag gagtcaacgt ttttttactg tgttcatcac aaaaccatta agctgagtca
     3661 tccatattgc aaacaataag gtagcaccgc ttccatcaaa acactagaac ataccgatgt
     3721 tttcacggat ttattggaaa caaatagcag atgagggcgt ttttacttgt aatgaaggcg
     3781 aatgagaagg caaggcttac aggcttgttg tttatgccaa gcatcaatac tcgctatgga
     3841 ggagcgccat cagcgataac aacaaagttt gcaagtgctt gcttggatat aaaaggcttg
     3901 acgcttaaga taaatagggc catacttgct tgcttttact atttttgttc ctcataagcc
     3961 tgattgacta cgctattata aacccctaca ttatcctata atcaacttca cttgtgtaac
     4021 gttgaatgag ttcaactgag taatactcgg tcggaaatat acgctccatc tggcaggaaa
     4081 cataggctgc atttatatct tgcacaatta tgcccttcgt aggtgtatgt tttcatgttt
     4141 tccattatgt caatgtagat gtatctttgg acaggcaact tttggatcaa taaagcacaa
     4201 tttttgagca atttgagata tccagtattt gtacgggagt cccaagattg tttagcctaa
     4261 aaggttgagg acttggatta ttcttccatc aagcgcctcc acccctcgca tcctatttct
     4321 tttgaagcat aggggacgag actgtctgta atttatggcg aagtatgaca agaaagattc
     4381 aaactttgtt ttacgtatgc cccaataaat tctatggtag cactactaga gacgtatttt
     4441 ggttttgtaa taattttcaa actgctcgct atgatttgtt acaagaaaca aacaaaagac
     4501 acgaccatag ggttcatgtt ggtggttgcg aagccacatc tttaaaacat tactgcaaat
     4561 tcagcaggag tgagtagcaa aatcgacaaa aactagagta cgcaacattc ccggtcgtaa
     4621 atttcttatc gtggcgccag aacattcccg tccaatgatg agaatgtctc taaaacaaat
     4681 gcagtcacag agattttctg cgacttctcg agtgagaatg ttgcttattg gtgctagtgg
     4741 cttgagatct tcgtcaagca aattggaatg ccccagattc tcaaacaagc acggcagaaa
     4801 ctgaaatacg gccaaaatta tgaacccttt tgtgggatct ttgtagaaat aggacctcag
     4861 gatagtatga tgaagaatgt tggcatgcaa gcttggcgta atcatggtca tagctgtttc
     4921 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4981 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5101 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7021 taggggttcc gcgcacattt ccccgaaaag tgccacctga gctttttctt tccaattttt
     7081 tttttttcgt cattatagaa atcattacga ccgagattcc cgggtaataa ctgatataat
     7141 taaattgaag ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt
     7201 tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca
     7261 ccctctacct tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca
     7321 gatcctgtag agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg
     7381 tcatctaaac ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc
     7441 atgtctcttt gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca
     7501 gtacccttag tatattctcc agtagctagg gagcccttgc atgacaattc tgctaacatc
     7561 aaaaggcctc taggttcctt tgttacttct tccgccgcct gcttcaaacc gctaacaata
     7621 cctgggccca ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca
     7681 cccgcagagt actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag
     7741 agtaaaaaat tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa
     7801 aaatcagtca agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca
     7861 actaactcca gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc
     7921 ttttcgtgca tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt
     7981 tccttatatg tagctttcga catgatttat cttcgtttcc tgcaggtttt tgttctgtgc
     8041 agttgggtta agaatactgg gcaatttcat gtttcttcaa caccacatat gcgtatatat
     8101 accaatctaa gtctgtgctc cttccttcgt tcttccttct gctcggagat taccgaatca
     8161 aaaaaatttc aaagaaaccg gaatcaaaaa aaagaacaaa aaaaaaaaag atgaattgaa
     8221 aagctctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8281 ccctttcgtc
//
