LOCUS       sequence-172606-        4840 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 4840)
  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 4840)
  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..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(286..305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(965..1756,1757..1825))
                     /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     1588..1607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1826..1930)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1998..2016
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2054..2076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2176..2195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2276..2294
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2276..2293
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      3041..3228
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3096..3120)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3303..3646
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4465..4662
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4769..4788)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4770..4788)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1981 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat
     2221 tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac atacgattta
     2281 ggtgacacta tagaacgcgg ccgccagctg aagcttcgta cgctgcaggt cgacggatcg
     2341 gtgacggtgc tggtttaatt aacggtggtg gttctatgtc taaaggtgaa gaattgttta
     2401 ctggtgttgt tccaattttg gttgaattgg atggtgatgt taatggtcat aaattttctg
     2461 ttagaggtga aggtgaaggt gatgctacta ttggtaaatt gactttgaaa tttatttgta
     2521 ctactggtaa attgccagtt ccatggccaa ctttggttac tactttgact tatggtgttc
     2581 aatgtttttc tagatatcca gatcatatga aacaacatga tttttttaaa tctgctatgc
     2641 cagaaggtta tgttcaagaa agaactattt cttttaaaga tgatggtaaa tataaaacta
     2701 gagctgttgt taaatttgaa ggtgatactt tggttaatag aattgaattg aaaggtactg
     2761 attttaaaga agatggtaat attttgggtc ataaattgga atataatttt aattctcata
     2821 atgtttatat tactgctgat aaacaaaaaa atggtattaa agctaatttt actgttagac
     2881 ataatgttga agatggttct gttcaattgg ctgatcatta tcaacaaaat actccaattg
     2941 gtgatggtcc agttttgttg ccagataatc attatttgtc tactcaaact gttttgtcta
     3001 aagatccaaa tgaaaaataa ggcgcgccac ttctaaataa gcgaatttct tatgatttat
     3061 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     3121 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     3181 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
     3241 ggataacagg gtaatataga tctgtttagc ttgcctcgtc cccgccgggt cacccggcca
     3301 gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat
     3361 gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca
     3421 tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cggacctgcg
     3481 agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga
     3541 gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa
     3601 tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatga cagtcaacac
     3661 taagacctat agtgagagag cagaaactca tgcctcacca gtagcacaac gattatttcg
     3721 attaatggaa ctgaagaaaa ccaatttatg tgcatcaatt gatgttgata ccactaagga
     3781 attccttgaa ttaattgata aattgggtcc ttatgtatgc ttaatcaaga ctcatattga
     3841 tataatcaat gatttttcct atgaatccac tattgaacca ttattagaac tttcacgtaa
     3901 acatcaattt atgatttttg aagatagaaa atttgctgat attggtaata ccgtgaagaa
     3961 acaatatatt ggtggagttt ataaaattag tagttgggca gatattacta atgctcatgg
     4021 tgtcactggg aatggagtag ttgaaggatt aaaacaggga gctaaagaaa ccaccaccaa
     4081 ccaagagcca agagggttat tgatgttagc tgaattatca tcagtgggat cattagcata
     4141 tggagaatat tctcaaaaaa ctgttgaaat tgctaaatcc gataaggaat ttgttattgg
     4201 atttattgcc caacgtgata tgggtggaca agaagaagga tttgattggc ttattatgac
     4261 acctggagtt ggattagatg ataaaggtga tggattagga caacaatata gaactgttga
     4321 tgaagttgtt agcactggaa ctgatattat cattgttggt agaggattgt ttggtaaagg
     4381 aagagatcca gatattgaag gtaaaaggta tagagatgct ggttggaatg cttatttgaa
     4441 aaagactggc caattataat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     4501 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     4561 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     4621 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     4681 cgccgccatc cagtgtcgaa aacgagctcg aattcatcga tgatatcaga tccactagtg
     4741 gcctatgcgg ccgcggatct gccggtctcc ctatagtgag tcgtattaat ttcgataagc
     4801 caggttaacc tgcattaatg aatcggccaa cgcgcgggga
//
