LOCUS       Exported                5073 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast integrative vector for expression of His-tagged POL30 (PCNA); 
            TRP1 marker.
ACCESSION   .
VERSION     .
KEYWORDS    YIp204-P30-His-POL30
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5073)
  AUTHORS   Davies AA, Ulrich HD
  TITLE     Detection of PCNA modifications in Saccharomyces cerevisiae.
  JOURNAL   Methods Mol Biol. 2012;920:543-67. doi: 
            10.1007/978-1-61779-998-3_36.
  PUBMED    22941627
REFERENCE   2  (bases 1 to 5073)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5073
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             576..593
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1404..1591
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1459..1483)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1615..1631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1615..1631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1628..1650)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1639..1655
                     /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)."
     promoter        complement(1663..1693)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1708..1729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1846..1863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2017..2605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2097..2116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2776..3636)
                     /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     3399..3418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3637..3741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3809..3827
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3829..3930
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             3931..4605
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(4724..4746)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4846..4865
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5059..8
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5073..17
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgccctta ttccttaact gctgaagaaa agcaacccta
       61 tgaatctaag gctcaagcag acaagaagag atacgaatct gaaaaggaat tgtacaatgc
      121 tacacgtgct tgatctaaca actcaatgtt ctctacctcc catacaacgt acttttgaaa
      181 cttcagaaag aagttgagta ggttgcatta tgtgttatta ttattactat tatcattatt
      241 ttacttatat atccttcttt tcttttatgt attctcttcc tttttcgcac aacttatgct
      301 gattcttcaa tatctaatta tttagcattt ttcttctcca tccgacgcac caaaacgcga
      361 aacgcgtaac tttttttttg gatttcaact gatagttttc gtactttgct tcctctggta
      421 cataaaatta tatataagaa acacttttgc tttagccttc ctttctttcc acttgcacct
      481 ttcactttcg ccgtcctttt tcactcacag caacaagcag caagcactaa gtacgcagtc
      541 aaaagagaga aaaaggatcc aatatgagag gatctcacca tcaccatcac catggcccgg
      601 gaaaaatgtt agaagcaaaa tttgaagaag catccctttt caagagaata attgatggtt
      661 tcaaagattg tgtccagttg gtcaatttcc aatgtaaaga agatggtatc attgcacaag
      721 ctgtcgatga ctcaagagtt ctattggtct ccttggaaat aggtgtcgaa gccttccaag
      781 aatatagatg tgaccatcct gttacgttag gtatggatct aacctcacta agtaaaatcc
      841 tacgttgtgg taacaacacc gatacattaa cactaattgc tgacaacaca ccggattcca
      901 tcatcttatt atttgaggat accaagaaag accgtatagc cgaatactct ctgaaattga
      961 tggatatcga tgctgatttc ttaaagattg aagaattaca gtacgactcc accctgtcat
     1021 tgccatcttc cgaattctct aaaattgttc gtgacttgtc ccaattgagt gattctatta
     1081 atatcatgat caccaaagaa acaataaagt ttgtagctga cggtgatatc ggatcaggtt
     1141 cagtcataat aaaaccattc gtggatatgg aacatcctga aacaagcatc aaacttgaaa
     1201 tggatcaacc tgtcgacttg acgttcggag ctaaatattt attggacatc attaagggct
     1261 cctccctttc tgatagagtt ggtatcaggc tctccagcga agctcctgct ttattccaat
     1321 ttgatttgaa gagtgggttc ctacagtttt tcttggctcc taaatttaat gacgaagaat
     1381 aactgcagcc aagctaattc cgggcgaatt tcttatgatt tatgattttt attattaaat
     1441 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     1501 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg
     1561 aggtcgctct tattgaccac acctctaccg gcatgcaagc ttggcgtaat catggtcata
     1621 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1681 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1741 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1801 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2581 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2641 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2701 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2761 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2821 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2881 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2941 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3001 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3061 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3121 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3181 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3241 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3301 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3361 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3421 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3481 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3541 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3601 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3661 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3721 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3781 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccaa ttcggtcgaa
     3841 aaaagaaaag gagagggcca agagggaggg cattggtgac tattgagcac gtgagtatac
     3901 gtgattaagc acacaaaggc agcttggagt atgtctgtta ttaatttcac aggtagttct
     3961 ggtccattgg tgaaagtttg cggcttgcag agcacagagg ccgcagaatg tgcactagat
     4021 tccgatgctg acttgctggg tattatatgt gtgcccaata gaaagagaac aattgacccg
     4081 gttattgcaa ggaaaatttc aagtcttgta aaagcatata aaaatagttc aggcactccg
     4141 aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg atgttttggc tctggtcaat
     4201 gattacggca ttgatatcgt ccaactgcat ggagatgagt cgtggcaaga ataccaagag
     4261 ttcctcggtt tgccagttat taaaagactc gtatttccaa aagactgcaa catactactc
     4321 agtgcagctt cacagaaacc tcattcgttt attcccttgt ttgattcaga agcaggtggg
     4381 acaggtgaac ttttggattg gaactcgatt tctgactggg ttggaaggca agagagcccc
     4441 gagagcttac attttatgtt agctggtgga ctgacgccag aaaatgttgg tgatgcgctt
     4501 agattaaatg gcgttattgg tgttgatgta agcggaggtg tggagacaaa tggtgtaaaa
     4561 gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga aataggttat tactgagtag
     4621 tatttattta agtattgttt gtgcacttgc ctgcaagcct tttgaaaagc aagcataaaa
     4681 gatcgccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4741 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4801 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     4861 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4921 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4981 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     5041 gggtaacgcc agggttttcc cagtcacgac gtt
//
