LOCUS       sequence-348222-        5414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Anp1-msGFP2 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5414)
  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 5414)
  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-ANP1-msGFP2
FEATURES             Location/Qualifiers
     source          1..5414
                     /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             631..1338
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(1912..1929)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1912..1928)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1921..1943)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2137..2156)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2256..2278
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2396..3199)
                     /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(3200..3415)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3422..3440)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3508..3612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3613..3681,3682..4473)
                     /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(3831..3850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4644..5232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5133..5152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5386..5403
                     /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 atgcacaaga tctgggagat tggatttgaa gatgtcagag accaatggaa cgaagaaaga
      301 gattcaatct taaaaaacat tgactccact ttgaacaata aagtcactgt agactggtct
      361 gaagagggag atggctcgga gctggtggac tcaaagggtg actttgtctc accaaacaac
      421 caacaacaac aacaacagca acaacaacaa caacagcagc agcagcaaca gcaacagcaa
      481 caacagctcg acggcaaccc gcaggggaaa cctcttgatg acaacgacaa gaacaagaaa
      541 aaacatccta aagaagttcc attagacttc gaccctgata gaaacttgga tccgggaagt
      601 tcggggtcca gcggttcttc aggcgctagt atggatagca ctgagagctt gttcacaggt
      661 gttgttccta tcttggtcga attagacggt gacgtaaacg gtcataagtt cagtgttaga
      721 ggtgaaggtg aaggtgacgc tactaacggt aaattgacat tgaagtttat ttgtactaca
      781 ggtaaattgc cagttccttg gccaactttg gttactacat tgacatacgg tgttcaatgt
      841 ttttcaagat acccagatca tatgaagcaa catgatttct ttaagtctgc tatgccagaa
      901 ggttacgttc aagaaagaac tatcactttt aaagatgatg gtacttacaa gacaagagca
      961 gaggttaagt tcgaaggtga cactttggtt aacagaatcg aattgaaggg tatcgatttt
     1021 aaagaagatg gtaatatttt gggtcataaa ttggaataca actacaactc acataacgtt
     1081 tacattacag ctgataaaca aaagaatggt attaaagcaa acttcaagat cagacataat
     1141 gttgaagatg gttctgttca attagcagat cattaccaac aaaacactcc aattggtgac
     1201 ggtccagttt tgttaccaga taaccattac ttgtctacac aatcaaaatt gtctaaagat
     1261 ccaaacgaaa agagagatca tatggttttg ttagaatttg ttactgctgc tggtatcaca
     1321 ggtggcagtg gcggttctta ataagcggcc gcgctcggtt ttttagtgac ctacatatga
     1381 agtacgaatt catatagttg tacatatata ccgataataa tatcaaacct ctattctttg
     1441 gtctttaact tttctttgag tgaattacaa taccattgtt atatcaatca ctatacacgg
     1501 ctcggtggcg aaattgaaat ttttttattt tcttctcttt cattatggct aggcaagaaa
     1561 tagcgacagc aaaacttaag aaaaaatgat agaacaaggt cccatccagt attgactagt
     1621 agctaaagct agaaggtaca cttaggtttt tctccctgtt tttaagtttg aaccattgat
     1681 aaggccgttc gcagacaagc ttgctaccat ccaattttcc ttcctctaaa gacttaataa
     1741 aggcatctaa ctccaccttt aaatcacaga tcacacaaag acaacataca atagaaaaat
     1801 ggttagctta acctttaaaa atttcaagaa ggaaaaagtt cctttagatc tggaaccttc
     1861 aaacacaatt ttagagacca agaccaagct tgcggtaccg agctcgaatt cactggccgt
     1921 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     1981 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2041 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     2101 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     2161 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2221 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2281 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcggg taataactga tataattaaa
     2341 ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt
     2401 tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct
     2461 ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc
     2521 ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat
     2581 ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt
     2641 ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac
     2701 ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa
     2761 ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg
     2821 ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg
     2881 cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta
     2941 aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat
     3001 cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta
     3061 actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt
     3121 cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct
     3181 tatatgtagc tttcgacatg atttatcttc gtttcctgca tgtttttgtt ctgtgcagtt
     3241 gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca
     3301 atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa
     3361 aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaaaagct
     3421 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3481 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3541 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3601 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3661 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3721 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3781 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3841 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3901 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3961 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4021 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4081 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4141 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4201 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4261 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4321 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4381 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4441 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4501 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4561 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4621 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4681 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4741 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4801 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4861 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4921 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4981 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5041 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5101 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5161 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5221 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5281 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5341 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5401 gaggaagcgg aaga
//
