LOCUS       sequence-348242-        7508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Sys1-msGFP3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7508)
  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 7508)
  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-SYS1-msGFPx3
FEATURES             Location/Qualifiers
     source          1..7508
                     /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(4006..4023)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4006..4022)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4015..4037)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4231..4250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4350..4372
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4490..5293)
                     /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(5294..5509)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5516..5534)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5602..5706
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5707..5775,5776..6567)
                     /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(5925..5944)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6738..7326
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7227..7246
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7480..7497
                     /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 ccaagcttct
      241 agaatcgaat ccagtgtggc attattacat ttacgacaac agaggtgttc aggactggaa
      301 atttgcagaa acgataaaac atctgctcga agcaatattt tcaatgttgt tgacgaactt
      361 ttacaaatgg ttaagaacgc aaaggattta tcaacttgag ttctgttcgt aaattatgta
      421 ccaccctgtt actcgtttca tattcacgct agagaagtac ctcagcttac tagatccatt
      481 cacacattgc aagtatacta aaactatata aaatttaagt acgtaaactc tttacttatt
      541 attatcttca atgtcatcac gtacttaatt atgttaaata tagattgtgt ataaataaga
      601 aataggatct gttttgaaaa attggccaaa aaggcagatt tattaagcct atagagataa
      661 ggttcttgcc atttttcata catttactgt tttctcttga atgactataa tattgccaaa
      721 attgcctcga atatgttgtt tcatctcctc gcggccgctt tacccgctgg agccggagct
      781 gccgtgagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
      841 gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac
      901 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
      961 ctccacgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt
     1021 gatatagacg ttgtggctgt tgaagttgta ctccagcttg tgccccagga tgttgccgtc
     1081 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     1141 cacctcggcg cgggtcttgt aggtgccgtc gtccttgaag gtgatggtgc gctcctggac
     1201 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     1261 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     1321 cttgccggtg gtgcagatga acttcagggt cagcttgccg ttggtggcat cgccctcgcc
     1381 ctcgccgcgc acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     1441 caccaccccg gtgaacaggc tctcagtgct atccatgcta gacccgctgg agccggagct
     1501 gccgtgagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     1561 gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac
     1621 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     1681 ctccacgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt
     1741 gatatagacg ttgtggctgt tgaagttgta ctccagcttg tgccccagga tgttgccgtc
     1801 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     1861 cacctcggcg cgggtcttgt aggtgccgtc gtccttgaag gtgatggtgc gctcctggac
     1921 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     1981 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     2041 cttgccggtg gtgcagatga acttcagggt cagcttgccg ttggtggcat cgccctcgcc
     2101 ctcgccgcgc acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     2161 caccaccccg gtgaacaggc tctcagtgct atccatgcta gacccgctgg agccggagct
     2221 gccgtgagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     2281 gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac
     2341 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     2401 ctccacgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt
     2461 gatatagacg ttgtggctgt tgaagttgta ctccagcttg tgccccagga tgttgccgtc
     2521 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     2581 cacctcggcg cgggtcttgt aggtgccgtc gtccttgaag gtgatggtgc gctcctggac
     2641 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     2701 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     2761 cttgccggtg gtgcagatga acttcagggt cagcttgccg ttggtggcat cgccctcgcc
     2821 ctcgccgcgc acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     2881 caccaccccg gtgaacaggc tctcagtgct atccatgcta gcgcctgaag aaccgctgga
     2941 ccccgaactt cctggatccc gtatttggct ttctaagtct tttagttgta ttggtgattg
     3001 ctccaattct gaatgattac tatgctgctg actaggcgta accaatccaa cttctccttc
     3061 attaggatca accaagccct caaaaaaggt gtctctaagc tctctccatc ttgttgtcca
     3121 tgtccccaaa aatattagta ttaaagatga taaaatttga agaaaaaacc aagaaaaaga
     3181 aggaaatttc cgagtataga ggaacaccac aataaaattg atggcatgta tagtgatagc
     3241 aaaatcccat gcgagtttac tccgtccaac gataacagtt aagaaaaaaa cacatattaa
     3301 tgaatccaga agccataaca gtgagattga taaaccgtat gcgttcgtaa agtcgatatt
     3361 ctcccaagaa aacagccatt ccttgatatt gagatcatat cccgctagct tcgcccaaca
     3421 ataaaaaaga acgatcgctg tggtgtaata aaaaatctgt aaaagaacaa tttgaagtcc
     3481 aatcttactt ggagagagag aatcttgttt gaagatctgg gaaggtttca actcattcgg
     3541 cactcgcaaa taccttctta tcgaaaccat gttacagtaa ttcaaatgcc tattttttat
     3601 ctcacttcta tgtgtaccct tccttagcct tcctaaagat catctcttaa catgaagtta
     3661 cagaatatta ccgtgaatat cgcgagctac gaatattttg ggtggggaga tatttattca
     3721 tgattagcct tgtcctatag tgttacaatg gagcaaattg ttcaccgtta gtattttgaa
     3781 tacgacaata ttaaatgaat aatagcgcct atctcttgtt acgtttggcg atttgtatac
     3841 cttgttaaat aaactgatta aaagccgggt tatggccgta cacaagttat tagaaagcaa
     3901 atacacttcc ttatatctgg tcacctagtt aataaaagga agtctttaat tttggcaaaa
     3961 accactatat atattagtca acaactgaaa tattaccagg aattcactgg ccgtcgtttt
     4021 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4081 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4141 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4201 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4261 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4321 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4381 gtcatcaccg aaacgcgcga gacgaaaggg cgggtaataa ctgatataat taaattgaag
     4441 ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct
     4501 ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct
     4561 tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag
     4621 agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac
     4681 ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt
     4741 gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag
     4801 tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc
     4861 taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca
     4921 ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt
     4981 actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat
     5041 tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca
     5101 agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca
     5161 gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca
     5221 tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg
     5281 tagctttcga catgatttat cttcgtttcc tgcatgtttt tgttctgtgc agttgggtta
     5341 agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa
     5401 gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc
     5461 aaggaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaaa agctggcctc
     5521 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5581 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5641 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5701 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5761 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5821 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5881 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5941 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6001 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6061 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6121 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6181 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6241 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6301 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6361 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6421 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6481 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6541 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6601 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6661 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6721 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6781 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6841 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6901 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6961 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7021 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7081 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7141 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7201 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7261 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7321 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7381 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7441 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7501 gcggaaga
//
