LOCUS       sequence-348217-        5754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Mnn9-msGFP2  in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5754)
  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 5754)
  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-MNN9-msGFP2
FEATURES             Location/Qualifiers
     source          1..5754
                     /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             856..1563
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(2252..2269)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2252..2268)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2261..2283)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2477..2496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2596..2618
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2736..3539)
                     /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(3540..3755)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3762..3780)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3848..3952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3953..4021,4022..4813)
                     /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(4171..4190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4984..5572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5473..5492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5726..5743
                     /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 atgcaggaaa gacgtgcagc aatggctttg gcgcgcaatg aattactatt ctccaccata
      301 ggacctcaca cttcttgggt gctgtggcta gatgccgata ttatagagac accaccatct
      361 ttaattcaag acatgaccaa acacaacaaa gctatcttag ctgcaaacat ttatcaaaga
      421 ttttacgatg aagagaagaa gcaaccatca atcagaccat acgatttcaa caactggcaa
      481 gaaagtgaca ccggtttaga aatagcctct cagatgggtg atgacgagat tattgtcgag
      541 ggttatgcag aaattgccac ttataggcca ttaatggctc atttctacga tgctaatggc
      601 gtaccaggtg aagagatggc gctggatggt gttggtggag gctgtacttt ggtcaaagca
      661 gaagttcaca gagacggtgc catgttccct aatttcccat tttatcactt gattgaaaca
      721 gaaggttttg ctaagatggc gaagagatta aactacgatg tatttggctt accaaactat
      781 ttggtttatc acatagagga agagaaccat tcggatccgg gaagttcggg gtccagcggt
      841 tcttcaggcg ctagtatgga tagcactgag agcttgttca caggtgttgt tcctatcttg
      901 gtcgaattag acggtgacgt aaacggtcat aagttcagtg ttagaggtga aggtgaaggt
      961 gacgctacta acggtaaatt gacattgaag tttatttgta ctacaggtaa attgccagtt
     1021 ccttggccaa ctttggttac tacattgaca tacggtgttc aatgtttttc aagataccca
     1081 gatcatatga agcaacatga tttctttaag tctgctatgc cagaaggtta cgttcaagaa
     1141 agaactatca cttttaaaga tgatggtact tacaagacaa gagcagaggt taagttcgaa
     1201 ggtgacactt tggttaacag aatcgaattg aagggtatcg attttaaaga agatggtaat
     1261 attttgggtc ataaattgga atacaactac aactcacata acgtttacat tacagctgat
     1321 aaacaaaaga atggtattaa agcaaacttc aagatcagac ataatgttga agatggttct
     1381 gttcaattag cagatcatta ccaacaaaac actccaattg gtgacggtcc agttttgtta
     1441 ccagataacc attacttgtc tacacaatca aaattgtcta aagatccaaa cgaaaagaga
     1501 gatcatatgg ttttgttaga atttgttact gctgctggta tcacaggtgg cagtggcggt
     1561 tcttaataag cggccgcgca actgagcaaa aagcatacag aagctatagc gttattgaaa
     1621 gataatattg ttgcgattat tagttgaaaa taaacaaaac aagatatata ttcatatata
     1681 tatatatata atttacacat gtatattaaa aaatgccgaa taaaaagtgc tacaacaact
     1741 aatttctgag cttgagaata aaccattcac attttatttc ttttggtgct tctttatcat
     1801 ttataaatcg gtggtcttga ctttttattc ctttctagtc taacaatcat ccattctata
     1861 gcgtctttaa caccttctcc agtcaatgcg cttattggta aaaccctact atctctagcg
     1921 cttatatgtt ccgcaatctt attaaatact tcttttatat cttgtacttc cattctatct
     1981 tgtctatctt gtttattggc cagcatcaag atgggtacac cttcaatttc ttcatccatt
     2041 acaactgact gtagggtcgt ggaacattcg tctaatcgtt ctctatctga actatccaca
     2101 atgaaaatta taccatgaca tagggaatag tattcggacc acattgatct cagtgattct
     2161 tgaccaccta catcccaaaa cttcaagatc tgtttactgt caacgggtat tgtcgccaca
     2221 ttttgtccta ccgggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact
     2281 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2341 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2401 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2461 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2521 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2581 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2641 gcgcgagacg aaagggcggg taataactga tataattaaa ttgaagctct aatttgtgag
     2701 tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc
     2761 aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc
     2821 tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc
     2881 acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc
     2941 ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg
     3001 ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta
     3061 gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt
     3121 acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca
     3181 ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact
     3241 gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat
     3301 aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt
     3361 gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg
     3421 gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc
     3481 ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg
     3541 atttatcttc gtttcctgca tgtttttgtt ctgtgcagtt gggttaagaa tactgggcaa
     3601 tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc
     3661 cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat
     3721 caaaaaaaag aataaaaaaa aaatgatgaa ttgaaaagct ggcctcgtga tacgcctatt
     3781 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3961 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4021 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4081 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4141 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4201 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4261 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4321 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4381 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4441 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4501 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4561 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4621 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4681 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4741 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4801 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5221 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5281 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5341 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5401 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5461 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5521 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5581 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5641 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5701 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga
//
