LOCUS       sequence-348245-        5523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Aur1-msGFP2 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5523)
  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 5523)
  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-AUR1-msGFP2
FEATURES             Location/Qualifiers
     source          1..5523
                     /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"
     CDS             802..1509
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(2021..2038)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2021..2037)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2030..2052)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2246..2265)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2365..2387
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2505..3308)
                     /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(3309..3524)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3531..3549)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3617..3721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3722..3790,3791..4582)
                     /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(3940..3959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4753..5341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5242..5261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5495..5512
                     /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 agaattgata agctactcgg tattaatatg tatactacag ctttttcaaa ttcctccgtc
      301 attttcggtg cttttccttc actgcattcc gggtgtgcta ctatggaagc cctgtttttc
      361 tgttattgtt ttccaaaatt gaagcccttg tttattgctt atgtttgctg gttatggtgg
      421 tcaactatgt atctgacaca ccattatttt gtagacctta tggcaggttc tgtgctgtca
      481 tacgttattt tccagtacac aaagtacaca catttaccaa ttgtagatac atctcttttt
      541 tgcagatggt catacacttc aattgagaaa tacgatatat caaagagtga tccattggct
      601 gcagattcaa acgatatcga aagtgtccct ttgtccaact tggaacttga ctttgatctt
      661 aatatgactg atgaacccag tgtaagccct tcgttatttg atggatctac ttctgtttct
      721 cgttcgtccg ccacgtctat aacgtcacta ggtgtaaaga gggctggtgg cagtggcggt
      781 tctggaggct caggagggag catggatagc actgagagct tgttcacagg tgttgttcct
      841 atcttggtcg aattagacgg tgacgtaaac ggtcataagt tcagtgttag aggtgaaggt
      901 gaaggtgacg ctactaacgg taaattgaca ttgaagttta tttgtactac aggtaaattg
      961 ccagttcctt ggccaacttt ggttactaca ttgacatacg gtgttcaatg tttttcaaga
     1021 tacccagatc atatgaagca acatgatttc tttaagtctg ctatgccaga aggttacgtt
     1081 caagaaagaa ctatcacttt taaagatgat ggtacttaca agacaagagc agaggttaag
     1141 ttcgaaggtg acactttggt taacagaatc gaattgaagg gtatcgattt taaagaagat
     1201 ggtaatattt tgggtcataa attggaatac aactacaact cacataacgt ttacattaca
     1261 gctgataaac aaaagaatgg tattaaagca aacttcaaga tcagacataa tgttgaagat
     1321 ggttctgttc aattagcaga tcattaccaa caaaacactc caattggtga cggtccagtt
     1381 ttgttaccag ataaccatta cttgtctaca caatcaaaat tgtctaaaga tccaaacgaa
     1441 aagagagatc atatggtttt gttagaattt gttactgctg ctggtatcac aggtggcagt
     1501 ggcggttctt aatgagtatt ttatctgcaa ttacggatac ggttggtctt atgtagatac
     1561 atataaatat atatcttttt ctttcttttt cttagtcagg attgtcgttt agcataatat
     1621 acatgtagtt tatttaatca cataccactg attatcttta gaattttata aatttttgaa
     1681 ataaatgggt ggcttttaat ggtgtctatg ttaagtgagg cttttagaat gctcttcctg
     1741 ctttgtttat tatatgtgta tgaaagatat gtatgtattt acatgtgttt gtagcgtccc
     1801 cagtcaaaac ctgtgcgcta tacctaaatg gattgataat cttcattcac taattctaaa
     1861 atagacttct tccccaaaga acggtgtaac gatgaggctc tatccagctg cttatctaaa
     1921 tcaactttaa cgatggatga tcttatgaca cggggatctt tctttaaagt tcttagaatt
     1981 tcagactgta ccgcagctga tgggtaccga gctcgaattc actggccgtc gttttacaac
     2041 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2101 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2161 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2221 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2281 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2341 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2401 caccgaaacg cgcgagacga aagggcgggt aataactgat ataattaaat tgaagctcta
     2461 atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg
     2521 catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca
     2581 tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc
     2641 acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca
     2701 ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca
     2761 ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat
     2821 tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt
     2881 tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca
     2941 ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc
     3001 aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac
     3061 ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata
     3121 tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat
     3181 tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata
     3241 ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct
     3301 ttcgacatga tttatcttcg tttcctgcat gtttttgttc tgtgcagttg ggttaagaat
     3361 actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg
     3421 tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga
     3481 aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagctg gcctcgtgat
     3541 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     3601 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3661 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3721 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3781 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3841 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3901 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3961 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4021 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4081 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4141 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4201 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4261 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4321 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4381 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4441 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4501 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4561 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4621 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4681 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4741 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4801 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4861 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4921 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4981 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5041 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5101 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5161 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5221 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5281 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5341 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5401 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5461 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5521 aga
//
