LOCUS       sequence-348213-        9408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iGFP-Drs2 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9408)
  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 9408)
  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-iGFP-DRS2
FEATURES             Location/Qualifiers
     source          1..9408
                     /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(910..938)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(5906..5923)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5906..5922)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5915..5937)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6131..6150)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6250..6272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6390..7193)
                     /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(7194..7409)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7416..7434)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7502..7606
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7607..7675,7676..8467)
                     /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(7825..7844)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8638..9226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9127..9146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9380..9397
                     /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 gtcgactcta gtgccaaaac taaacgtcct aaagtcgaaa tcgaatacga
      301 ggaggaacac gaagttcaaa atgctgagca agaagtggca caataagctc acgcactata
      361 ctacgcaaca ttacatgatt tcagtttcta ttattgaatg aacaggattt ttccacgtgg
      421 attgggcagc gcactgtacc ataataacaa ccagttataa aatatttcaa ttcttttaca
      481 tatataaaaa tacataaaga ctattatttt tgtaagatgg cccattctgt aacacttaaa
      541 tttcttatca aaaaaaaaaa aaataatacc tttagtagct gttgggctaa taacattgca
      601 aattagaagc ttactattag aagcttacta ttacgggatt attgaggtaa aggttatttg
      661 aagcgtaaat cactgaaatc ttcctgatca cgtgattggc tttttttgcc agaaaaccca
      721 aatctgttta atgccactaa gttgaagaaa catataatca acaattgttg gcaattctaa
      781 ggggaatagt gcaacaagaa gccagtaacg ccaaatagag gttagcgcca agcaataatt
      841 tactggtagg ttctttcatt ccctgatttt accatgaata tggatagcac tgagagcctt
      901 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      961 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     1021 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt cacttatggt
     1081 gttcagtgct tttcaagata cccggatcat atgaaacggc atgacttttt caagagtgcc
     1141 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     1201 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     1261 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     1321 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     1381 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     1441 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     1501 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     1561 gggattacac atggcagctc cggctccagc gggtccggaa gtggtagctc agggtctagt
     1621 ggatccgacg acagagaaac ccccccaaag aggaaacctg gggaggacga cacgcttttc
     1681 gatattgact tcttagacga cacgacttct cattctggat cccgttcaaa ggtcaccaac
     1741 tcacatgcga acgcgaatta tataccacca agtcatgtac ttcccgaaga gactatcgac
     1801 ttagacgctg atgatgataa cattgaaaat gatgtccatg aaaatctatt tatgagcaat
     1861 aaccatgacg accaaacgtc ttggaatgcc aatagattcg actcagatgc atatcaacca
     1921 caatccctac gagcagtcaa gcctcccggt ttatttgcta ggtttggtaa tggtctcaaa
     1981 aatgccttta cgttcaaacg gaagaaaggg cccgaaagtt tcgaaatgaa tcattacaat
     2041 gcggtgacta ataacgaatt agacgataat tatctagact cgaggaataa gttcaacatt
     2101 aaaattcttt tcaaccggta cattttaaga aagaacgttg gcgacgcaga gggcaatggt
     2161 gaaccaagag tcatccacat caacgattct ctcgctaatt cctccttcgg ttatagtgac
     2221 aaccatatat ctaccaccaa gtataatttt gcgacctttc tgcctaaatt tttgttccaa
     2281 gaattttcca aatacgctaa tctgtttttt ttatgtacat ctgccattca acaggtgcct
     2341 cacgtctcgc caactaatag atacaccaca attggtactt tattagtggt tttgattgtt
     2401 tctgccatga aggaatgtat cgaagatatc aagagagcta attctgataa agaattaaat
     2461 aattcgacag cagaaatatt ttcagaagca catgatgact ttgttgagaa acgatggatt
     2521 gatattcgtg taggtgacat aattagagta aaatcagagg aacctattcc tgctgatacc
     2581 ataattttgt catcttcgga accggagggt ctttgctaca ttgaaactgc caacttggat
     2641 ggtgaaacaa atttgaaaat caaacagtcg agagtagaaa ctgccaaatt tatagacgtt
     2701 aaaactttga agaacatgaa cggaaaagtt gtttccgaac agccgaactc tagcttgtat
     2761 acttatgaag gtacaatgac tttaaatgat cgtcaaatac cgttatctcc tgaccaaatg
     2821 attttaagag gtgcaacttt aagaaatact gcatggatat ttggtttagt tatcttcact
     2881 ggtcatgaaa ctaagttatt gcgtaatgct actgcaaccc caattaaaag aaccgcggtt
     2941 gagaaaatta tcaacagaca gattattgca ttgttcacag ttttaatcgt gctaatttta
     3001 atttcttcaa ttggtaatgt tattatgtct actgcagatg ccaaacattt atcgtacctt
     3061 tacctggagg gtaccaacaa ggctggctta ttcttcaaag actttttaac attttggatt
     3121 ctattttcga atctagttcc tatttctcta tttgtcaccg ttgaattaat caaatattat
     3181 caggctttta tgataggttc agatctagat ttgtactacg aaaaaactga taccccaact
     3241 gtggttcgta catcctcttt ggttgaagaa cttggtcaaa tagaatatat attcagtgac
     3301 aagacaggaa ctttaacaag aaatattatg gaatttaaat cctgctctat tgcaggccat
     3361 tgctatattg ataaaatacc tgaagataaa acggccactg ttgaagatgg gattgaagtt
     3421 ggttatagaa aatttgacga tttgaagaaa aagttaaatg atccctctga tgaggattca
     3481 cctattatca atgacttctt aacattactg gctacctgtc atactgtcat tccagaattt
     3541 caaagcgatg gatctattaa atatcaagca gcctctccag atgaaggtgc cctcgttcaa
     3601 ggtggtgcag atttagggta taagtttatc atccgtaaac caaactctgt aactgtttta
     3661 ttggaggaaa ctggcgagga aaaagagtat caactactta acatttgtga atttaattcc
     3721 accaggaaga gaatgagcgc tatatttaga tttccggatg gttcgataaa gttattctgt
     3781 aaaggtgctg atactgtcat tctggaaaga ttggatgatg aagctaatca atatgtagaa
     3841 gctacaatga gacatttaga agattatgca tctgagggtt tgcggacgtt gtgtttggcg
     3901 atgagagata tctctgaagg agaatatgaa gaatggaata gcatttataa tgaggctgcc
     3961 acaacattag ataacagagc cgagaagcta gacgaagccg caaatctaat cgagaaaaat
     4021 ctaatattaa ttggtgcaac tgctattgaa gataagttac aggacggtgt tccagaaact
     4081 atccacacat tacaagaagc gggtattaaa atttgggttt taactggtga cagacaggaa
     4141 actgctatta acattggtat gagttgccgt ttattgagtg aagacatgaa tttgttgatc
     4201 atcaacgagg aaaccagaga tgatactgag agaaatttgt tagagaagat taacgctcta
     4261 aacgagcatc aattgtcaac acatgatatg aataccttag cgctcgtcat tgatgggaag
     4321 tcattaggct ttgccttgga accagaatta gaagattatt tgctaactgt ggcgaagctt
     4381 tgtaaagcgg ttatatgttg ccgtgtatct ccactacaga aagccttggt tgttaaaatg
     4441 gtaaaaagaa agtcgtcttc actactgcta gccattggcg atggtgccaa cgatgttagt
     4501 atgatacagg cagctcatgt tggtgtcggt attagtggta tggaaggtat gcaagcggct
     4561 cgttcagctg atatagctgt tggccaattt aaatttttaa aaaaactatt acttgtccat
     4621 ggctcctggt cttatcaaag aatttctgcc gcaattttgt actcttttta caagaataca
     4681 gcattataca tgacgcagtt ttggtacgtt tttgccaatg ccttttcagg tcaatccatt
     4741 atggaatcat ggacaatgag tttctacaac ttattcttca ctgtttggcc cccttttgtc
     4801 attggtgtat ttgatcaatt tgttagtagt agattacttg agcggtaccc acaattgtac
     4861 aaattaggac agaagggtca atttttctct gtttacatct tctggggatg gattattaat
     4921 ggctttttcc attctgcaat agtatttatt ggcaccatat tgatttacag atatgggttt
     4981 gccttaaata tgcacggcga actagctgat cattggtctt ggggtgtaac tgtctacact
     5041 acaagtgtca ttattgtttt gggaaaggct gctttggtaa ctaatcaatg gacgaaattc
     5101 accttaattg ccattcctgg ttccctctta ttttggttaa tattcttccc aatttatgct
     5161 tctatatttc ctcatgctaa catctcaaga gagtattatg gtgtggttaa acacacgtat
     5221 ggatccggtg tattttggtt aactttgatc gttttaccaa tttttgcact ggtaagagat
     5281 tttctatgga agtactataa aagaatgtat gaaccagaaa cgtatcatgt tattcaagaa
     5341 atgcagaaat acaatatcag cgactctagg ccgcacgttc agcaatttca aaatgccatc
     5401 aggaaggtga ggcaagtgca aagaatgaaa aaacaaagag gatttgcctt ttcacaggct
     5461 gaagagggtg ggcaagaaaa aattgtcaga atgtatgata ctactcaaaa gagaggtaag
     5521 tacggtgaat tacaagatgc ttcggcgaac ccctttaatg acaataatgg actgggaagt
     5581 aacgactttg aaagtgcaga accgttcatt gaaaatccat ttgctgatgg taatcaaaat
     5641 tcaaatagat tcagttcttc gagagatgat atttcatttg atatatgagg cacagaaaca
     5701 aagagcagaa gacaaaaaat acaaaaataa aaataagata aaaagaaaag tcattattac
     5761 cattactgaa atatgtacat ttaccataaa atatatccgt tatatagaag ttataatatt
     5821 aagccatcac tttgttatcg gtttttcatt cctcctgggg ttgcttcgac tgggtacgta
     5881 ccttatgtct tctaggctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     5941 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     6001 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     6061 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     6121 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     6181 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     6241 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     6301 gacgaaaggg cgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     6361 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     6421 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     6481 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     6541 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     6601 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     6661 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     6721 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     6781 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     6841 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     6901 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     6961 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     7021 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     7081 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     7141 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     7201 cttcgtttcc tgcatgtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     7261 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     7321 tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa
     7381 aaagaataaa aaaaaaatga tgaattgaaa agctggcctc gtgatacgcc tatttttata
     7441 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     7501 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     7561 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7621 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7681 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7741 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7801 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     7861 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7921 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7981 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     8041 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     8101 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     8161 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     8221 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     8281 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     8341 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     8401 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     8461 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8521 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8581 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     8641 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     8701 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     8761 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     8821 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     8881 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     8941 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     9001 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     9061 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     9121 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     9181 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     9241 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     9301 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     9361 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaga
//
