LOCUS       sequence-169646-        4838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal PCR tagging module for yeast codon optimized split 
            GFP1-10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4838)
  AUTHORS   Smoyer CJ, Katta SS, Gardner JM, Stoltz L, McCroskey S, Bradford WD,
            McClain M, Smith SE, Slaughter BD, Unruh JR, Jaspersen SL
  TITLE     Analysis of membrane proteins localizing to the inner nuclear 
            envelope in living cells.
  JOURNAL   J Cell Biol. 2016 Nov 21;215(4):575-590. Epub 2016 Nov 9.
  PUBMED    27831485
REFERENCE   2  (bases 1 to 4838)
  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: "2016-11-21"; volume: "215(4)"; pages: "575-590. Epub 
            2016 Nov 9"
COMMENT     Sequence Label: pSJ1256 (pFA6-link-yGFP1-10-CaURA3MX)
FEATURES             Location/Qualifiers
     source          1..4838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..80)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(62..80)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(167..184)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(338..926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(418..437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1097..1888,1889..1957))
                     /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     1720..1739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1958..2062)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2130..2148
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2186..2208)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2308..2327
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2408..2426
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2408..2425
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      3172..3359
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3227..3251)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3434..3777
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4596..4793
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct
       61 ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc
      121 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
      181 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      241 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      301 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      361 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      421 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      481 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
      541 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      601 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      661 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      721 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
      781 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      841 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      901 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      961 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1021 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1081 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1141 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1201 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1261 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1321 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1381 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1441 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1501 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1561 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1621 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1681 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1741 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1801 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1861 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1921 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1981 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2041 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     2101 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     2161 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2221 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2281 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2341 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
     2401 acatacgatt taggtgacac tatagaacgc ggccgccagc tgaagcttcg tacgctgcag
     2461 gtcgacggat cggtgacggt gctggttaat taacggtggt ggttctatgt ctaaaggtga
     2521 agaattgttt actggtgttg ttccaatttt ggttgaattg gatggtgatg ttaatggtca
     2581 taaattttct gttagaggtg aaggtgaagg tgatgctact attggtaaat tgactttgaa
     2641 atttatttgt actactggta aattgccagt tccatggcca actttggtta ctactttgac
     2701 ttatggtgtt caatgttttt ctagatatcc agatcatatg aaacaacatg atttttttaa
     2761 atctgctatg ccagaaggtt atgttcaaga aagaactatt tcttttaaag atgatggtaa
     2821 atataaaact agagctgttg ttaaatttga aggtgatact ttggttaata gaattgaatt
     2881 gaaaggtact gattttaaag aagatggtaa tattttgggt cataaattgg aatataattt
     2941 taattctcat aatgtttata ttactgctga taaacaaaaa aatggtatta aagctaattt
     3001 tactgttaga cataatgttg aagatggttc tgttcaattg gctgatcatt atcaacaaaa
     3061 tactccaatt ggtgatggtc cagttttgtt gccagataat cattatttgt ctactcaaac
     3121 tgttttgtct aaagatccaa atgaaaaata aggcgcgcca cttctaaata agcgaatttc
     3181 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     3241 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
     3301 gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc
     3361 agatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt ccccgccggg
     3421 tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag
     3481 ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc
     3541 atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct
     3601 gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc
     3661 gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact
     3721 tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg
     3781 acagtcaaca ctaagaccta tagtgagaga gcagaaactc atgcctcacc agtagcacag
     3841 cgattatttc gattaatgga actgaagaaa accaatttat gtgcatcaat tgacgttgat
     3901 accactaagg aattccttga attaattgat aaattaggtc cttatgtatg cttaatcaag
     3961 actcatattg atataatcaa tgatttttcc tatgaatcca ctattgaacc attattagaa
     4021 ctttcacgta aacatcaatt tatgattttt gaagatagaa aatttgctga tattggtaat
     4081 accgtaaaga aacaatatat tggtggagtt tataaaatta gtagttgggc agatattacc
     4141 aatgctcatg gtgtcactgg gaatggagtg gttgaaggat taaaacaggg agctaaagaa
     4201 accaccacca accaagagcc aagagggtta ttgatgttag ctgaattatc atcagtggga
     4261 tcattagcat atggagaata ttctcaaaaa actgttgaaa ttgctaaatc cgataaggaa
     4321 tttgttattg gatttattgc ccaacgtgat atgggtggcc aagaagaagg atttgattgg
     4381 cttattatga cacctggagt tggattagat gataaaggtg atggattagg acaacaatat
     4441 agaactgttg atgaagttgt tagcactgga actgatatta tcattgttgg tagaggattg
     4501 tttggtaaag gaagagatcc agatattgaa ggtaaaaggt atagaaatgc tggttggaat
     4561 gcttatttga aaaagactgg ccaattataa tcagtactga caataaaaag attcttgttt
     4621 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     4681 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     4741 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga
     4801 ttcgatacta acgccgccat ccagtgtcga aaacgagc
//
