LOCUS       sequence-11858-e        5737 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 5737)
  AUTHORS   Rossanese OW, Soderholm J, Bevis BJ, Sears IB, O'Connor J, 
            Williamson EK, Glick BS
  TITLE     Golgi structure correlates with transitional endoplasmic reticulum 
            organization in Pichia pastoris and Saccharomyces cerevisiae.
  JOURNAL   J Cell Biol. 1999 Apr 5. 145(1):69-81.
  PUBMED    10189369
REFERENCE   2  (bases 1 to 5737)
  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. 1999 Apr 5. 145(1):69-81."
COMMENT     Sequence Label: pUC19-URA3-3'SEC13-GFP
FEATURES             Location/Qualifiers
     source          1..5737
                     /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      884..893
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(890..892,893..895,896..1609)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(890..892,893..895,896..1606)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(935..956)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1196..1215)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1543..1564
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2350..2366)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2350..2366)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2363..2385)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2374..2390
                     /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(2398..2404,2405..2422,2423..2428))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2443..2464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2581..2598)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2752..3340)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2832..2851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3511..4302,4303..4371))
                     /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     4134..4153
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4529..5332)
                     /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(5333..5549)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     5710..5728
                     /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 cgagctcggt acccggggat
      421 caactctgct tcttgggctc cagctaccat cgaagaagat ggtgaacaca acggtactaa
      481 agaatctcgc aagtttgtta ctgggggtgc tgacaatttg gtaaagattt ggaagtacaa
      541 ttcagatgcc caaacttatg ttctggaaag caccttagaa ggtcacagcg attgggttag
      601 agacgtagca tggtcaccta ctgttcttct acgttcttat ttggccagtg tttctcaaga
      661 tcgcacctgt attatttgga ctcaagacaa tgaacaaggc ccatggaaaa aaactttatt
      721 aaaagaagaa aaattcccag atgttttatg gagagccagt tggtctttgt caggtaatgt
      781 actagctctt tccggtggcg ataataaagt tactttatgg aaggaaaatc ttgagggtaa
      841 atgggaaccc gctggtgaag ttcatcagta cgatccaccg gtcgccacca tggtgagcaa
      901 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      961 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1021 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1081 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1141 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1201 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1261 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1321 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1381 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1441 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1501 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1561 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgtag
     1621 tatcaagaat ttaaaatgaa acatctcaaa agaaaaaaga atgcaaatga gtctatcgac
     1681 gaatttgaag ggaaataaac gcataatgta cagtaacgta taacaattaa agatttgtgg
     1741 aagttttcaa aaacttttca acttttttct ttgttttttt tttgcaactt cttatattaa
     1801 tattgtcata gatatttctt atacaaaaac aagcgaacaa aaataatcga cgtatataca
     1861 atagatatat aagactgttt ttcttcaata gaacaggcga aattattcta ccggccgaag
     1921 gtacatcttc ccgctatgta ataaatagag gtatttaagt tataacaatt ataattcccc
     1981 cattacggca gcaattaagt tttgcacgtc attagaggcg attttaccat tctccagtac
     2041 ttcagcgtga gtcgctttgc ctttttctaa gggtacaggt gattcgtcca acgcactggc
     2101 agggctatcc accacgcaag tattggtaat caaacttaag gccaaaaccc tccatccgca
     2161 atgtcttgca acaatgactt cgggaacagt actcattccg acagcatctc ctcccaacat
     2221 ccttatcatt ttggattctg ctcttgtttc gaaagtgggt ccagatacaa aagtataagt
     2281 accttcatgc agtggccttt gaatcttgag ctctttccat ttcttaaata aaagcttggc
     2341 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2401 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2461 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2521 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2581 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2641 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2701 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2761 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2821 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2881 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2941 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3001 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3061 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3121 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3181 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3241 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3301 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3361 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3421 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3481 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3541 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3601 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3661 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3721 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3781 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3841 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3901 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3961 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4021 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4081 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4141 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4201 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4261 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4321 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4381 ttttcaatag ctttttcttt ccaatttttt ttttttcgtc attatagaaa tcattacgac
     4441 cgagattccc gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
     4501 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
     4561 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
     4621 atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc
     4681 tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca
     4741 accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa
     4801 aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca gtagctaggg
     4861 agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt
     4921 ccgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa
     4981 tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac
     5041 caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct
     5101 ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta
     5161 gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa
     5221 catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag
     5281 caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc
     5341 ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg caatttcatg
     5401 tttcttcaac accacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
     5461 cttccttctg ctcggagatt accgaatcaa aaaaatttca aagaaaccgg aatcaaaaaa
     5521 aagaacaaaa aaaaaaaaga tgaattgaaa agctattatt gaagcattta tcagggttat
     5581 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5641 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5701 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
