LOCUS       pUSE-URA3-Sec7-m        6817 bp DNA     circular SYN 17-JAN-2024
DEFINITION  Plasmid for pop-in/pop-out replacement of Sec7 with a Sec7-msYFP2 
            fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6817)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported May 6, 2024 from SnapGene 7.1.0b
            https://www.snapgene.com
COMMENT     Linearize with BsiWI for integration at the SEC7 locus.
FEATURES             Location/Qualifiers
     source          1..6817
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             397..975
                     /codon_start=1
                     /label=SEC7
                     /translation="NSIVVKCVLQLLMIELLNELFENEDFAHCIPYKEAIRITRLLEKS
                     YEFSRDFNEDYGLRTRLVEARVVDKIPNLLKQETSAAAVLLDIMFQLYLNDDEKKADLI
                     TRLITICIQVVEGYVSLDDRTMERSINAWRSVIVEILQGYYEFDDEDFRLYCPAMYALV
                     IQILDKSVPTELRHAIKQFLSRVGELYLST"
     CDS             994..1701
                     /codon_start=1
                     /label=msYFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     misc_feature    1713..3414
                     /label=3' SEC7
     rep_origin      complement(3836..4424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4595..5386,5387..5455))
                     /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"
     promoter        complement(5456..5560)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(5727..6530)
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6531..6747)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ctattgttgt caaatgtgtt
      421 ttacagcttt tgatgataga gctgctcaac gaattattcg agaacgaaga ttttgcccac
      481 tgtatccctt acaaggaagc cattagaatt acaagattgt tggagaaatc gtacgaattt
      541 tctcgtgact ttaatgaaga ttatgggtta agaacaagac tagtagaggc tcgtgtagtt
      601 gataaaatac caaacctgtt gaaacaagaa acgagtgctg ctgcagttct ccttgacatt
      661 atgtttcagt tgtatctgaa cgatgatgag aagaaggctg atttaataac tcgtctgatc
      721 accatttgca tccaggtcgt agagggttat gtttctttgg atgatagaac catggaacgt
      781 agtatcaatg cctggcgttc agtgatagtt gaaatattac aaggttacta cgaatttgat
      841 gatgaagact ttagactata ctgtcctgct atgtatgctc tggtgattca aatattagat
      901 aagagcgttc caacggagtt gagacatgct ataaaacaat ttctaagcag agttggtgaa
      961 ttataccttt ctacggatcc actagtcgcc accatggata gcactgagag cctgttcacc
     1021 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1081 cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc
     1141 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacct ccctgggcta cggcctgcag
     1201 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1261 gaaggctacg tccaggagcg caccatcacc ttcaaggacg acggcaccta caagacccgc
     1321 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1381 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa cagccacaac
     1441 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1501 aacgtggagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1561 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1621 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1681 actggaggca gtggcggttc ttaaagcggc cgggctttgt tgaatggtcc agcagattaa
     1741 aatatgctta gttgtagaac acagaaaaaa tgagaaggtt gatatacttt tactcatatt
     1801 aatcctgatt tagtggcggt gatccttttt agattcgttt aattcataaa caaggtacat
     1861 catatagcaa taacaaatat gtaaactatg tagaaagact tatcaagttt agccattttg
     1921 gtcctcattc atgggcattt tatactaact tcatttttgc cgaaataaga aaggatgtaa
     1981 tcttaaccta aaatatgtaa gcaattaagg gcgcacgtag tcacttgcga acaaatagga
     2041 aaagaaagaa acaagcacgg atactgaatg actgagtgta ttactacacg atttatctca
     2101 acgaatttaa gtgtggagga cgatcggctg cacgtaggag tcaacgtttt tttactgtgt
     2161 tcatcacaaa accattaagc tgagtcatcc atattgcaaa caataaggta gcaccgcttc
     2221 catcaaaaca ctagaacata ccgatgtttt cacggattta ttggaaacaa atagcagatg
     2281 agggcgtttt tacttgtaat gaaggcgaat gagaaggcaa ggcttacagg cttgttgttt
     2341 atgccaagca tcaatactcg ctatggagga gcgccatcag cgataacaac aaagtttgca
     2401 agtgcttgct tggatataaa aggcttgacg cttaagataa atagggccat acttgcttgc
     2461 ttttactatt tttgttcctc ataagcctga ttgactacgc tattataaac ccctacatta
     2521 tcctataatc aacttcactt gtgtaacgtt gaatgagttc aactgagtaa tactcggtcg
     2581 gaaatatacg ctccatctgg caggaaacat aggctgcatt tatatcttgc acaattatgc
     2641 ccttcgtagg tgtatgtttt catgttttcc attatgtcaa tgtagatgta tctttggaca
     2701 ggcaactttt ggatcaataa agcacaattt ttgagcaatt tgagatatcc agtatttgta
     2761 cgggagtccc aagattgttt agcctaaaag gttgaggact tggattattc ttccatcaag
     2821 cgcctccacc cctcgcatcc tatttctttt gaagcatagg ggacgagact gtctgtaatt
     2881 tatggcgaag tatgacaaga aagattcaaa ctttgtttta cgtatgcccc aataaattct
     2941 atggtagcac tactagagac gtattttggt tttgtaataa ttttcaaact gctcgctatg
     3001 atttgttaca agaaacaaac aaaagacacg accatagggt tcatgttggt ggttgcgaag
     3061 ccacatcttt aaaacattac tgcaaattca gcaggagtga gtagcaaaat cgacaaaaac
     3121 tagagtacgc aacattcccg gtcgtaaatt tcttatcgtg gcgccagaac attcccgtcc
     3181 aatgatgaga atgtctctaa aacaaatgca gtcacagaga ttttctgcga cttctcgagt
     3241 gagaatgttg cttattggtg ctagtggctt gagatcttcg tcaagcaaat tggaatgccc
     3301 cagattctca aacaagcacg gcagaaactg aaatacggcc aaaattatga acccttttgt
     3361 gggatctttg tagaaatagg acctcaggat agtatgatga agaatgttgg catgcaagct
     3421 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3481 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3541 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3601 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4261 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4381 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4441 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4501 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4561 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4621 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4681 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4741 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4801 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4861 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4921 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4981 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5041 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5101 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5161 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5221 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5281 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5341 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5401 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5461 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5521 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5581 cacctgagct ttttctttcc aatttttttt ttttcgtcat tatagaaatc attacgaccg
     5641 agattcccgg gtaataactg atataattaa attgaagctc taatttgtga gtttagtata
     5701 catgcattta cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct
     5761 tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat
     5821 agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta
     5881 tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac
     5941 caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa
     6001 tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt agctagggag
     6061 cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttcc
     6121 gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg
     6181 tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca
     6241 atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt
     6301 agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt
     6361 aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca
     6421 tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca
     6481 acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt
     6541 cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt
     6601 tcttcaacac cacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct
     6661 tccttctgct cggagattac cgaatcaaaa aaatttcaaa gaaaccggaa tcaaaaaaaa
     6721 gaacaaaaaa aaaaaagatg aattgaaaag ctctaagaaa ccattattat catgacatta
     6781 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
