LOCUS       sequence-172609-        5262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal PCR tagging cassette for yeast codon optimized split 
            GFP1-10 with CDC42 promoter and KAR2 signal sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5262)
  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 5262)
  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: pSJ1726 (pFA6-NATMX-CDC42pr-KAR2SS-GFP1-10)
FEATURES             Location/Qualifiers
     source          1..5262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(184..527)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        complement(640..658)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(641..658)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(739..758)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     858..880
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(918..936)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1004..1108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1109..1177,1178..1969)
                     /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     complement(1327..1346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2140..2728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2629..2648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2882..2899
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2986..3005
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2986..3004
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(join(4670..5262,1..527))
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     terminator      complement(4670..4867)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4873..5262,1..183))
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
ORIGIN
        1 ggggaacacc ttggtcaggg gcgggtccac cggcacctcc cgcagggtga agccgtcccc
       61 ggtggcggtg acgcggaaga cggtgtcggt ggtgaaggac ccatccagtg cctcgatggc
      121 ctcggcgtcc cccgggacac tggtgcggta ccggtaagcc gtgtcgtcaa gagtggtacc
      181 catggttgtt tatgttcgga tgtgatgtga gaactgtatc ctagcaagat tttaaaagga
      241 agtatatgaa agaagaacct cagtggcaaa tcctaacctt ttatatttct ctacaggggc
      301 gcggcgtggg gacaattcaa cgcgtctgtg aggggagcgt ttccctgctc gcaggtctgc
      361 agcgaggagc cgtaattttt gcttcgcgcc gtgcggccat caaaatgtat ggatgcaaat
      421 gattatacat ggggatgtat gggctaaatg tacgggcgac agtcacatca tgcccctgag
      481 ctgcgcacgt caagactgtc aaggagggta ttctgggcct ccatgtcgct ggccgggtga
      541 cccggcgggg acgaggcaag ctaaacagat ctggcgcgcc ttaattaacc cggggatccg
      601 tcgacctgca gcgtacgaag cttcagctgg cggccgcgtt ctatagtgtc acctaaatcg
      661 tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg acaatatgtc
      721 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
      781 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
      841 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
      901 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
      961 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1021 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1081 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1141 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1201 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1261 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1321 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1381 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1441 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1501 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1561 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1621 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1681 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1741 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1801 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1861 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1921 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1981 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2041 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2101 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2161 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2221 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2281 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2341 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2401 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2461 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2521 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2581 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2641 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2701 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2761 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2821 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2881 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     2941 gcgttggccg attcattaat gcaggttaac ctggcttatc gaaattaata cgactcacta
     3001 tagggagacc ggcagatccg cggccgcata ggccactagt ggatctgata tcatcgatga
     3061 gaaccaccac cagaaccacc acctttttca tttggatctt tagacaaaac agtttgagta
     3121 gacaaataat gattatctgg caacaaaact ggaccatcac caattggagt attttgttga
     3181 taatgatcag ccaattgaac agaaccatct tcaacattat gtctaacagt aaaattagct
     3241 ttaataccat tcttttgttt atcagcagta atataaacat tatgagaatt aaaattatat
     3301 tccaatttat gacccaaaat attaccatct tctttaaaat cagtaccttt caattcaatt
     3361 ctattaacca aagtatcacc ttcaaattta acaacagctc tagttttata tttaccatca
     3421 tctttaaaag aaatagttct ttcttgaaca taaccttctg gcatagcaga tttaaaaaaa
     3481 tcatgttgtt tcatatgatc tggatatcta gaaaaacatt gaacaccata agtcaaagta
     3541 gtaaccaaag ttggccatgg aactggcaat ttaccagtag tacaaataaa tttcaaagtc
     3601 aatttaccaa tagtagcatc accttcacct tcacctctaa cagaaaattt atgaccatta
     3661 acatcaccat ccaattcaac caaaattgga acaacaccag taaacaattc ttcaccttta
     3721 gacatggata aaggtaatat taccacgaaa agggcgtaca ggaccacgga gagtggtacc
     3781 agcagcttgc cagcgcttag tctgttgaaa aacatggtat gtttgatacg ctttttccct
     3841 tgagactact cttggggact ttaaaaaagt aaaacagttc ctttgtggaa gagctaatac
     3901 gtttatttct tgttttataa gttttgcttc tctaattcgt ttgttaacct atctcctcag
     3961 gaaaatagaa tagtataata aataacgaat ataatttttt gcgacaaagg aaacctattg
     4021 caaaaagtga agaatgacag agaaggataa tcagtatatc tcggtacacc tggaacctat
     4081 ttatcaaaac gctgctcttg ccttttccaa gaatcgcgtc tatcatcgat caatcaacct
     4141 tgtttgcttt tttttatcac tttttcccgt atcttgtttc gtttggaagg gcccaatgga
     4201 agcagtatta cccgtattga atgacccttg aatcgacggt ggaagacatc tttaatgcga
     4261 tgtttagaag acaatgtgcc gtatttacct gtatatgtgg attatctaac agtctacaat
     4321 acaaaattag gtgtaacgta gctacaacag ttgatagaaa taatgcaact ttttttaaaa
     4381 aaaaaagaga aagtgctcga ggtggggatt gaacccacga cggtcgcgtt gcttttaaag
     4441 gcctgtttga aaggtctttg gcacagaaac ttcggaaacc gaatgttgct ataagcacga
     4501 agctctaacc actgagctac acgagcattt tcgaaagaga cgtaaattaa attgttattt
     4561 ataccgtaga tgattgtttt tcacgaaatt acccttttga gttccggcct gtctagagcg
     4621 gcgagctcgt tttcgacact ggatggcggc gttagtatcg aatcgacagc agtatagcga
     4681 ccagcattca catacgattg acgcatgata ttactttctg cgcacttaac ttcgcatctg
     4741 ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct aacatttgat taaaatagaa
     4801 caactacaat ataaaaaaac tatacaaatg acaagttctt gaaaacaaga atctttttat
     4861 tgtcagtact gattaggggc agggcatgct catgtagagc gcctgctcgc cgtccgaggc
     4921 ggtgccgtcg tacagggcgg tgtccaggcc gcagagggtg aaccccatcc gccggtacgc
     4981 gtggatcgcc ggtgcgttga cgttggtgac ctccagccag aggtgcccgg cgccccgctc
     5041 gcgggcgaac tccgtcgcga gccccatcaa cgcgcgcccg accccgtgcc cccggtgctc
     5101 cggggcgacc tcgatgtcct cgacggtcag ccggcggttc cagccggagt acgagacgac
     5161 cacgaagccc gccaggtcgc cgtcgtcccc gtacgcgacg aacgtccggg agtccgggtc
     5221 gccgtcctcc ccgtcgtccg attcgtcgtc cgattcgtcg tc
//
