LOCUS       sequence-348209-        6967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Sec2-msGFPx3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6967)
  AUTHORS   Casler JC, Johnson N, Krahn AH, Pantazopoulou A, Day KJ, Glick BS
  TITLE     Clathrin adaptors mediate two sequential pathways of intra-Golgi 
            recycling.
  JOURNAL   J Cell Biol. 2022 Jan 3;221(1). pii: 212747. doi: 
            10.1083/jcb.202103199. Epub 2021 Nov 5.
  PUBMED    34739034
REFERENCE   2  (bases 1 to 6967)
  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."; date: "2022-01-3"; volume: "221(1). pii: 212747. doi"; 
            pages: " 10.1083/jcb.202103199. Epub 2021 Nov 5"
COMMENT     Sequence Label: YIplac211-SEC2-msGFPx3
FEATURES             Location/Qualifiers
     source          1..6967
                     /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"
     primer_bind     complement(3465..3482)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3465..3481)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3474..3496)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3690..3709)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3809..3831
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3949..4752)
                     /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(4753..4968)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4975..4993)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5061..5165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5166..5234,5235..6026)
                     /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(5384..5403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6197..6785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6686..6705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6939..6956
                     /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 atgcctgcag gacaaagctc tttggaagca ggacgcaaaa agaccttttt cttcatctag
      301 cgcagaagaa agtcaaaaga gcgacgcttt tgactttgaa agtggagata tggaaaatga
      361 aattacgggt gaatcttcaa gtgatgaatc ctcttcagat gggtcttcca cagataattc
      421 aaccgcagat agttcatcag aagatgaatc ttctctagca gattctacta caagtagcgc
      481 agattctagt tcaccagaga gtatagacaa cggtgaaggt gatgacacgg taacgaaaga
      541 tgacaaaagt agcattaagt cggcaaataa taatgaagaa aacagcgatt gcggagataa
      601 aaaaggtcgt tctataataa agaaaaaggc tcctcaacgc aagatacaaa agaagaagtt
      661 actccaagat ttggatgatt tagaagagca atttagagaa gaaagcgcaa ttgatcagac
      721 tgaatttgaa aacgctgagt ctaatgtaaa acaaaatata tcctcgaaga gggcatcttc
      781 tggagacgag aactctaaga aggataataa tgaaaaaact ttaaagacaa acttgacgat
      841 aggcgacaaa acccaagaac aaatagggga aaatagtcct agtagcggat tgcatgcgtc
      901 ttcaagcaat gatgacaatt tcgatgatgc ccaggaacag caaacggatc caggaagttc
      961 ggggtccagc ggttcttcag gcgctagcat ggatagcact gagagcctgt tcaccggggt
     1021 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg
     1081 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
     1141 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1201 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1261 ctacgtccag gagcgcacca tcaccttcaa ggacgacggc acctacaaga cccgcgccga
     1321 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1381 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacgtcta
     1441 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacgt
     1501 ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1561 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     1621 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca
     1681 cggcagctcc ggctccagcg ggtctagcat ggatagcact gagagcctgt tcaccggggt
     1741 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg
     1801 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
     1861 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1921 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1981 ctacgtccag gagcgcacca tcaccttcaa ggacgacggc acctacaaga cccgcgccga
     2041 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2101 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacgtcta
     2161 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacgt
     2221 ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2281 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     2341 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca
     2401 cggcagctcc ggctccagcg ggtctagcat ggatagcact gagagcctgt tcaccggggt
     2461 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg
     2521 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
     2581 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     2641 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2701 ctacgtccag gagcgcacca tcaccttcaa ggacgacggc acctacaaga cccgcgccga
     2761 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2821 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacgtcta
     2881 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacgt
     2941 ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3001 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     3061 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca
     3121 cggcagctcc ggctccagcg ggtaaagcgg ccgcattata taatatttaa ttttatgtgt
     3181 cctaatggac atttgagttc cttaagatgt tcaataactt agaaaaatat aggtaatgat
     3241 aaagaatgaa tcgcatgtat tgttttaata ttatgtcaat actttgaact taaaaacaga
     3301 gtaaagaagt ggttctaaaa ttacacgtat taaagggatt aattactaca tattcattct
     3361 caatcatcac tatcaccttg gctcaaaaca actctagact ttttgccaaa aaggcggatg
     3421 gcctcttcat cttcatcgct ttcatcggta ccgagctcga attcactggc cgtcgtttta
     3481 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3541 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3601 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3661 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3721 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3781 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3841 tcatcaccga aacgcgcgag acgaaagggc gggtaataac tgatataatt aaattgaagc
     3901 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     3961 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     4021 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     4081 gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc
     4141 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     4201 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     4261 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     4321 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     4381 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     4441 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     4501 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     4561 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     4621 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     4681 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     4741 agctttcgac atgatttatc ttcgtttcct gcatgttttt gttctgtgca gttgggttaa
     4801 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     4861 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     4921 aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa gctggcctcg
     4981 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5041 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5101 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5161 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5221 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5281 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5341 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5401 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5461 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5521 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5581 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5641 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5701 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5761 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5821 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5881 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5941 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6001 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6061 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6121 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6181 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6301 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6361 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6421 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6481 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6541 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6601 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6661 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6721 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6781 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6841 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6901 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6961 cggaaga
//
