LOCUS       sequence-348220-        5474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Stv1-iGFP in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5474)
  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 5474)
  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-STV1-iGFP
FEATURES             Location/Qualifiers
     source          1..5474
                     /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(770..798)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(1972..1989)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1972..1988)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1981..2003)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2197..2216)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2316..2338
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2456..3259)
                     /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(3260..3475)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3482..3500)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3568..3672
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3673..3741,3742..4533)
                     /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(3891..3910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4704..5292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5193..5212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5446..5463
                     /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 atgcgggatg atcatcagtg atgttgccag tgttgcagac agtatcaatg aaagcgttgg
      301 tggaggtgaa caggggccat ttaatttcgg cgatgtcatg attcatcagg taatccatac
      361 tattgaattc tgtttaaatt gtatctctca tacagcatca tatctacgtc tttgggcact
      421 atcgttggcc catgcgcaat tatctagtgt tttatgggat atgacaattt caaacgcttt
      481 cagttctaaa aactcagggt cgcctttggc tgtcatgaag gtggttttct tatttgctat
      541 gtggtttgtt ttaaccgttt gtatcctagt tttcatggaa ggtacttccg caatgttaca
      601 tgcgctacgt ttgcattggg tggaggcaat gtccaaattt tttgaaggtg aaggatatgc
      661 gtacgagcca ttttcatttc gtgcaataat agaagcggat ccaggaagtt cggggtccag
      721 cggttcttca ggcagttcca tggatagcac tgagagcctt ttcactggag ttgtcccaat
      781 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
      841 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
      901 tgttccttgg ccaacacttg tcactacttt aacttatggt gttcagtgct tttcaagata
      961 cccggatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     1021 ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt
     1081 tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg
     1141 aaacattctt gggcacaaat tggaatacaa ctataactca cacaatgtat acatcatggc
     1201 agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca ttgaagatgg
     1261 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     1321 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     1381 gagagaccac atggtcctta aagagtttgt aacagctgct gggattacac atggcagctc
     1441 cggctccagc gggtaaagcg gccgctaacc ttaaattact gtaaattttc tcatctaccc
     1501 actgcttttt tcgttttatt tttttttaca catacagaca tataaacgtt gtagaattaa
     1561 acgataaaat ataatatttc ttcacatagt ttacggtata tatatgtctg ggtcggtcaa
     1621 gtggtctacc aataggtcat aaatttgagc agggatctta gcgatgaacc cgacgcccag
     1681 ccgaatgatc tcgtcagcat caaaatccgg aaactgtcgt agtaaattaa caggggaatc
     1741 cgatttgaaa accttggatc tcattttaac taggaatgcc accacaaaga gaatattcat
     1801 atggaatcca tatgcaaaca tgaaatccca cagtttaatg acctgatcaa gcggtttatt
     1861 gcagctggat aacgtgagta tggaaggcat accataaatc tctgctgtaa gtaaattgtc
     1921 cgatagaaat ttactcagct ttggatcaat gattctgaga gaaatcaatt cactggccgt
     1981 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     2041 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2101 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     2161 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     2221 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2281 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2341 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcggg taataactga tataattaaa
     2401 ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt
     2461 tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct
     2521 ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc
     2581 ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat
     2641 ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt
     2701 ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac
     2761 ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa
     2821 ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg
     2881 ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg
     2941 cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta
     3001 aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat
     3061 cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta
     3121 actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt
     3181 cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct
     3241 tatatgtagc tttcgacatg atttatcttc gtttcctgca tgtttttgtt ctgtgcagtt
     3301 gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca
     3361 atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa
     3421 aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaaaagct
     3481 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3541 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3601 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3661 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3721 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3781 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3841 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3901 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3961 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4021 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4081 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4141 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4201 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4261 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4321 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4381 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4441 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4501 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4561 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4621 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4681 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4741 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4801 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4861 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4921 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4981 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5041 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5101 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5161 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5221 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5281 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5341 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5401 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5461 gaggaagcgg aaga
//
