LOCUS       sequence-348253-        9240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Abp1-iGFPx6 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9240)
  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 9240)
  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-ABP1-iGFPx6
FEATURES             Location/Qualifiers
     source          1..9240
                     /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(5738..5755)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5738..5754)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5747..5769)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5963..5982)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6082..6104
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6222..7025)
                     /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(7026..7241)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7248..7266)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7334..7438
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7439..7507,7508..8299)
                     /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(7657..7676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8470..9058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8959..8978
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9212..9229
                     /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 atgccacaag aacctgtgaa agctgaagaa gccgagcagc ctaagactga ttacaagaag
      301 atcggcaacc cattacccgg tatgcacatt gaagcggata atgaggaaga accagaagag
      361 aatgatgatg actgggatga tgatgaagac gaggctgctc aacctccttt gccttcgagg
      421 aatgttgcgt caggagcacc agtgcaaaaa gaagagcctg aacaagaaga gatcgcccca
      481 agcttacctt ctagaaactc gatcccagct ccaaaacaag aagaagcacc tgaacaagca
      541 cctgaagaag aaattgaaga agaagctgag gaagccgctc cacagctgcc atcaagaagc
      601 tctgcagctc ctcctccgcc tccaagacga gcaactccag agaaaaagcc aaaggaaaat
      661 ccttgggcca cagcagaata tgattacgat gctgcagaag ataacgaact gacctttgtg
      721 gaaaatgaca agattatcaa tattgaattt gtcgacgatg actggtggct aggggaacta
      781 gagaaagacg gctcaaaagg tctcttcccc agcaattatg tgtctttggg caacacggat
      841 ccaggaagtt cggggtccag cggttcttca ggcgctagca tggatagcac tgagagcctt
      901 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      961 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     1021 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     1081 gttcagtgct tttcaagata cccggatcat atgaaacagc 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 gggtctagca tggatagcac tgagagcctt
     1621 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     1681 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     1741 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     1801 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     1861 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     1921 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     1981 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     2041 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     2101 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     2161 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     2221 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     2281 gggattacac atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt
     2341 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     2401 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     2461 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     2521 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     2581 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     2641 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     2701 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     2761 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     2821 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     2881 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     2941 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     3001 gggattacac atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt
     3061 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     3121 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     3181 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     3241 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     3301 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     3361 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     3421 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     3481 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     3541 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     3601 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     3661 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     3721 gggattacac atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt
     3781 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     3841 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     3901 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     3961 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     4021 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     4081 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     4141 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     4201 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     4261 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     4321 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     4381 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     4441 gggattacac atggcagctc cggctccagc gggtctagca tggatagcac tgagagcctt
     4501 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     4561 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     4621 tgcactactg gaaaactacc tgttccttgg ccaacacttg tcactacttt aacttatggt
     4681 gttcagtgct tttcaagata cccggatcat atgaaacagc atgacttttt caagagtgcc
     4741 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     4801 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     4861 attgatttta aagaagatgg aaacattctt gggcacaaat tggaatacaa ctataactca
     4921 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     4981 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     5041 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     5101 tcgaaagatc ccaacgaaaa gagagaccac atggtcctta aagagtttgt aacagctgct
     5161 gggattacac atggcagctc cggctccagc gggtaaagcg gccgcaatca cacgtcagcg
     5221 tcatgctatt atattattct tacgtaaaaa aatacttaca gagacgttaa agcttttgta
     5281 tagatacttt cacgagcctt ccctttcagt aaaatgaaca aattattttg ccttgtcctt
     5341 ctttgtttca agcgtttata tgactcctct acagtattat ttttcaacag acggtgtgcc
     5401 cgtacggcgt tgtcaaaacc tcgaacataa aacaacactg aaacaaactt attgcactcc
     5461 tgcatcgcat acagatctga tacttactac aaccataatg ctgttttaga atcgcatcag
     5521 gtgtttacca gaagtgtatg gtacatggtg ctttccttat gctcactatt tcggatttac
     5581 ttctagtcta tgtatagatt tgttagagac cagttttaag ctacaatgga acgttaagat
     5641 taagaaatgc atgaagagtc tcccagtaga tatataaagc tgaattaaat tcagtacaat
     5701 ggagtttcca cgtctttctg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc
     5761 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     5821 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5881 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5941 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     6001 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     6061 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     6121 cgaaacgcgc gagacgaaag ggcgggtaat aactgatata attaaattga agctctaatt
     6181 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     6241 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     6301 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     6361 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     6421 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     6481 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     6541 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     6601 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     6661 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     6721 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     6781 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     6841 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     6901 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     6961 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     7021 gacatgattt atcttcgttt cctgcatgtt tttgttctgt gcagttgggt taagaatact
     7081 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     7141 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     7201 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaagctggcc tcgtgatacg
     7261 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     7321 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     7381 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     7441 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     7501 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     7561 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     7621 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     7681 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     7741 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7801 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7861 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7921 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7981 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     8041 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     8101 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     8161 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     8221 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     8281 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     8341 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     8401 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     8461 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     8521 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     8581 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     8641 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     8701 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     8761 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8821 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8881 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8941 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     9001 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     9061 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     9121 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     9181 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
//
