LOCUS       sequence-375783-        5896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single-integration vector for integration of MCP-GFPEnvy at the URA3
            locus in Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5896)
  AUTHORS   Brouwer I, de Kort MAC, Lenstra TL
  TITLE     Measuring Transcription Dynamics of Individual Genes Inside Living 
            Cells.
  JOURNAL   Methods Mol Biol. 2024;2694:235-265. doi: 
            10.1007/978-1-0716-3377-9_12.
  PUBMED    37824008
REFERENCE   2  (bases 1 to 5896)
  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: "Methods Mol
            Biol."; date: "2024"; volume: "2694:235-265. doi"; pages: " 
            10.1007/978-1-0716-3377-9_12"
COMMENT     Sequence Label: SIVURA MCP-GFPenvy
FEATURES             Location/Qualifiers
     source          1..5896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     256..273
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     257..273
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(302..318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(302..318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    319..352
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        389..732
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1551..1748
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1772..1805
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1806..1825
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1806..1824
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2843..2863
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      3614..3861
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3630..3648)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3884..3902)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     rep_origin      complement(4190..4778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4270..4289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4949..5740,5741..5809))
                     /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     5572..5591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5810..5873)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taacgtgatg atatttcata aataatgtaa ttctatatat gttaattacc ttttttgcga
       61 ggcatattta tggtgaagga taagttttga ccatcaaaga aggttaatgt ggctgtggtt
      121 tcagggtcca taatgctttt caattcatct tttttttttt tgttcttttt tttgattccg
      181 gtttctttga aatttttttg attcggtaat ctccgagcag aaggaagaac gaaggaagga
      241 gcacagactt agatttgtaa aacgacggcc agtgcatgcg tatcggacgt ctcccttcat
      301 ggtcatagct gtttcctgat aacttcgtat aatgtatgct atacgaagtt attagcttgc
      361 ctcgtccccg ccgggtcacc cggcgagcga catggaggcc cagaataccc tccttgacag
      421 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
      481 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
      541 ttacggctcc tcgctccaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
      601 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
      661 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
      721 acataaacaa ccatgacagt caacactaat acctatagtg agagagcaga aactcatgcc
      781 tcaccagtag cacagcgatt atttcgatta atggaactga agaaaaccaa tttatgtgca
      841 tcaattgacg ttgataccac taaggagttc cttgaattaa ttgataaatt aggtccttat
      901 gtatgcttaa tcaagactca tattgatata atcaatgatt tttcctatga atccactatt
      961 gaaccattat tagaactttc acgtaaacat caatttatga tttttgaaga tagaaaattt
     1021 gctgatattg gtaataccgt aaagaaacaa tatattggtg gagtttataa aattagtagt
     1081 tgggcagata ttaccaatgc tcatggtgtc actgggaatg gagtggttga aggattaaaa
     1141 cagggagcta aagaaaccac caccaaccaa gagccaagag ggttattgat gttagctgaa
     1201 ttatcatcag tgggatcatt agcatacgga gaatattctc aaaaaactgt tgaaattgct
     1261 aaatccgata aggaatttgt tattggattt attgcccaac gtgatatggg tggccaagaa
     1321 gaaggatttg attggcttat tatgacacct ggagttggat tagatgataa aggtgatgga
     1381 ttaggacaac aatatagaac tgttgatgaa gttgttagca ctggaactga tattatcatt
     1441 gttggtagag gattgtttgg taaaggaaga gatccagata ttgaaggtaa aaggtataga
     1501 aatgctggtt ggaatgctta tttgaaaaag actggccaat tataatcagt actgacaata
     1561 aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc
     1621 tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag
     1681 atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg
     1741 ctatactgct gtcgattcga tactaacgcc gataacttcg tataatgtat gctatacgaa
     1801 gttattaata cgactcacta tagggcgaat tggagctcgt attcaagata ttcttatcaa
     1861 agcacttatt ttttcattat atttctatat tgatttaagg acggtacgct attttctttg
     1921 taccttttac tatgcataaa aaaaggggaa gtgtttaaca atcgacactt acttaatatg
     1981 ttttgccgcc cttcataaga gggtgtttct aaaatttatt gggcaagaat gagatggact
     2041 cgcaccctac atgacgttta aatatttagt gttaaggttc agaacatgca ccaggtgcga
     2101 catgtgttgc gattatcatg acaatgtctc tatccgagat gcatttgtag tatcaattga
     2161 tgcgtattat gacatgattt acatagcata catcgtcaaa catgatatta tattcttttt
     2221 ttgataaatg tacggattta aagctgtcga atatattttc tgaaatttct tggagctgac
     2281 gcaaaatttt caaaggtgct aaaatttttc aagatttctc acttttgctt ggtaacaaag
     2341 aatgatggca ttgcattttt accaccggta catttaactg ctatttctca cgtttctttc
     2401 cctatcctta agtaattctt ttacaatcta agaaaaccac gatcaaacaa ataaatcagc
     2461 atctagaatg ctagccgtta aaatggcttc taactttact cagttcgttc tcgtcgacaa
     2521 tggcggaact ggcgacgtga ctgtcgcccc aagcaacttc gctaacggga tcgctgaatg
     2581 gatcagctct aactcgcgtt cacaggctta caaagtaacc tgtagcgttc gtcagagctc
     2641 tgcgcagaat cgcaaataca ccatcaaagt cgaggtgcct aaaggcgcct ggcgttcgta
     2701 cttaaatatg gaactaacca ttccaatttt cgccacgaat tccgactgcg agcttattgt
     2761 taaggcaatg caaggtctcc taaaagatgg aaacccgatt ccctcagcaa tcgcagcaaa
     2821 ctccggcatc tacgcggatc caccaaaaaa gaagagaaag gtaaaaccgg tcgccaccat
     2881 catgtctaaa ggcgaggaat tgtttacagg tgtagttcct attttggtcg aactagatgg
     2941 agatgtcaac ggtcataagt tttctgtacg tggtgaaggc gaaggtgatg ctactaatgg
     3001 caaactgact ctgaagttca tctgtacgac cggaaaattg ccagttccat ggcctacatt
     3061 agtaacaaca ttatgttacg gagttcaatg ctttgctaga tacccagatc acatgaagca
     3121 gcatgacttt ttcaaaagtg ctatgccaga aggttatgtg caggagagaa ctatctcctt
     3181 caaagatgat ggtacttata agacaagagc agaggtaaag tttgaaggcg atacattggt
     3241 taatagaatt gagcttaaag gcattgattt taaggaagat gggaatatcc tcggacataa
     3301 acttgaatac aatttcaatt cacataatgt ttatatcaca gcagataaac aaaagaatgg
     3361 tataaaagcg aactttaaga caagacataa cgtggaagat ggtagcgtcc aattagccga
     3421 ccattaccaa caaaacactc ctatcgggga cgggccagtt ctccttccag acaaccacta
     3481 cctatcaacc caatctgttc tatctaaaga ccctaatgaa aaacgagatc acatggtctt
     3541 gttagagttc gtgactgccg ctggtataac attaggaatg gacgaattgt acaaataagg
     3601 atcctaactc gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     3661 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     3721 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     3781 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     3841 cgctcgaagg ctttaatttg cggccggtac ccagcttttg ttccctttag tgagggttaa
     3901 tttcctgaaa actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc
     3961 aatttaatta tatcagttat tacgcgggaa tctcggtcgt aatgatttct ataatgacga
     4021 aaaaaaaaaa attggaaaga aaatgcttca tggcctttat aaaaaggaac tatccaatac
     4081 ctcgccagaa ccaagtaaca gtattttacg gggcacaaat caagaacaat aagacaggac
     4141 tgtaaagatg gacgcattta attaattcga aggccgcgtt gctggcgttt ttccataggc
     4201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4621 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4921 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4981 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5041 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5101 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5161 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5221 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5281 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5341 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5401 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5461 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5521 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5581 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5641 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5701 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5761 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5821 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaag
     5881 tatcgttcga attaat
//
