LOCUS       sequence-169650-        8968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luminal split GFP11 reporter with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8968)
  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 8968)
  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: pSJ1602 (pRS315-NOP1pr-mCherry-SCS2TM-GFP11)
FEATURES             Location/Qualifiers
     source          1..8968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..58)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(42..58)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(91..97,98..115,116..121))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    136..157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        223..598
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     promoter        790..1005
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             complement(1569..2663)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMSKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTAEVGDAVAEEVKKILA"
     promoter        complement(2664..3071)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(3857..3874)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4856..5444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4936..4955)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5615..6406,6407..6475))
                     /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     6238..6257
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6476..6580)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6648..6666
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6704..6726)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7039..7061
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7053..7070
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7054..7070
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7101..7117
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7101..7117
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             8556..8603
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
ORIGIN
        1 ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
       61 tgaaattgtt atcccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      121 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      181 tttccagtcg ggaaacctgt cgtgccagca gatctgaatt aattcttgaa taatacataa
      241 cttttcttaa aagaatcaaa gacagataaa atttaagaga tattaaatat tagtgagaag
      301 ccgagaattt tgtaacacca acataacact gacatcttta acaactttta attatgatac
      361 atttcttacg tcatgattga ttattacagc tatgctgaca aatgactctt gttgcatggc
      421 tacgaaccgg gtaatactaa gtgattgact cttgctgacc ttttattaag aactaaatgg
      481 acaatattat ggagcatttc atgtataaat tggtgcgtaa aatcgttgga tctctcttct
      541 aagtacatcc tactataaca atcaagaaaa acaagaaaat cggacaaaac aatcaagtat
      601 ggattctaga acagttggta tattaggagg gggacaattg ggacgtatga ttgttgaggc
      661 agcaaacagg ctcaacatta agacggtaat actagatgct gaaaattctc ctgccaaaca
      721 aataagcaac tccaatgacc acgttaatgc atcagagcag attgtactga gagtgcacca
      781 taccaccttt tcaattcatc attttttttt tattcttttt tttgatttcg gtttccttga
      841 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
      901 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
      961 acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc gaaagctaca
     1021 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaaggcaga ttgtactgag
     1081 agtgcaccat atcgactacg tcgttaaggc cgtttctgac agagtaaaat tcttgaggga
     1141 actttcacca ttatgggaaa tggttcaaga aggtattgac ttaaactcca tcaaatggtc
     1201 aggtcattga gtgtttttta tttgttgtat tttttttttt ttagagaaaa tcctccaata
     1261 tataaattag gaatcatagt ttcatgattt tctgttacac ctaacttttt gtgtggtgcc
     1321 ctcctccttg tcaatattaa tgttaaagtg caattctttt tccttatcac gttgagccat
     1381 tagtatcaat ttgcttacct gtattccttt acatcctcct ttttctcctt cttgataaat
     1441 gtatgtagat tgcgtatata gtttcgtcta ccctatgaac atattccatt ttgtaatttc
     1501 gtgtcgtttc tattatgaat ttcatttata aagtttatgt acaaatatca taaaaaaaga
     1561 gaatcttttt aagcaaggat tttcttaact tcttcggcga cagcatcacc gacttcggcg
     1621 gtactgttgg aaccacctaa atcaccagtt ctgatacctg catccaaaac ctttttaact
     1681 gcatcttcaa tggccttacc ttcttcaggc aagttcaatg acaatttcga catcattgca
     1741 gcagacaaga tagtggcgat agggttgacc ttattctttg gcaaatctgg agcagaaccg
     1801 tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc caaggacgca
     1861 gatggcaaca aacccaagga acctgggata acggaggctt catcggagat gatatcacca
     1921 aacatgttgc tggtgattat aataccattt aggtgggttg ggttcttaac taggatcatg
     1981 gcggcagaat caatcaattg atgttgaacc ttcaatgtag ggaattcgtt cttgatggtt
     2041 tcctccacag tttttctcca taatcttgaa gaggccaaaa cattagcttt atccaaggac
     2101 caaataggca atggtggctc atgttgtagg gccatgaaag cggccattct tgtgattctt
     2161 tgcacttctg gaacggtgta ttgttcacta tcccaagcga caccatcacc atcgtcttcc
     2221 tttctcttac caaagtaaat acctcccact aattctctga caacaacgaa gtcagtacct
     2281 ttagcaaatt gtggcttgat tggagataag tctaaaagag agtcggatgc aaagttacat
     2341 ggtcttaagt tggcgtacaa ttgaagttct ttacggattt ttagtaaacc ttgttcaggt
     2401 ctaacactac cggtacccca tttaggacca cccacagcac ctaacaaaac ggcatcagcc
     2461 ttcttggagg cttccagcgc ctcatctgga agtggaacac ctgtagcatc gatagcagca
     2521 ccaccaatta aatgattttc gaaatcgaac ttgacattgg aacgaacatc agaaatagct
     2581 ttaagaacct taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc tggcaaaacg
     2641 acgatcttct taggggcaga cattagaatg gtatatcctt gaaatatata tatatatatt
     2701 gctgaaatgt aaaaggtaag aaaagttaga aagtaagacg attgctaacc acctattgga
     2761 aaaaacaata ggtccttaaa taatattgtc aacttcaagt attgtgatgc aagcatttag
     2821 tcatgaacgc ttctctattc tatatgaaaa gccggttccg gcgctctcac ctttcctttt
     2881 tctcccaatt tttcagttga aaaaggtata tgcgtcaggc gacctctgaa attaacaaaa
     2941 aatttccagt catcgaattt gattctgtgc gatagcgccc ctgtgtgttc tcgttatgtt
     3001 gaggaaaaaa ataatggttg ctaagagatt cgaactcttg catcttacga tacctgagta
     3061 ttcccacagt taactgcggt caagatattt cttgaatcag gcgccttaga ccgctcggcc
     3121 aaacaaccaa ttacttgttg agaaatagag tataattatc ctataaatat aacgcttttg
     3181 aacacacatg aacagggaag tacaggacaa ttgattttga agagaatgtg gattttgatg
     3241 taattgttgg gattccattt ttaataaggc aataatatta ggtatgtaga tatactagaa
     3301 gttctcctcg accggtcgat atgcggtgtg aaataccgca cagagggtga acgttacaga
     3361 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     3421 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     3481 ttaccctatg cggtgtgaaa taccgcacag atgcgtagga gattcaagtt atacaacgaa
     3541 aatggaacag tttttattaa agcaagaaga agaagttctt gtcaaagcac aaaagttaga
     3601 aactgtcggt tacgaagctt atctagaaaa caagtaatat ataagtttat tgatatactt
     3661 gtacagcaaa taattataaa atgatatacc tattttttag gctttgttat gattacatca
     3721 aatgtggact tcatacatag aaatcaacgc ttacaggtgt ccttttttaa gaatttcata
     3781 cataagatct gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3841 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3901 cggtatcagc atcgatgcta gagtcgaatg gaaattacgt actttcaacg caagagcaag
     3961 acaaaagaaa gcctaaaact gtcttttcgt gtcggtcgtc caatatcatc gtaaacgtgt
     4021 gtagagcctt aaattctaat ttctaacttt aaattttttt tgctattata atactaattt
     4081 ctaactttca acctatttta catcttcgga aaacaaaaac tattttttct ttaatttctt
     4141 tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt ataattattt
     4201 ttatagcacg tgatgaaaag aaatatagtc ttcaaggtta tattatacag gtttatatat
     4261 tattttactt atttttgttg ttctttttct tggagtattt agcattaaca acttcgaccc
     4321 atcgcgatga cgaaattgca cgtttaaagg agaagtcgga atcgttgcaa gaagaaataa
     4381 attagtttga cgttttattc cttttcttct gcacagaagt gtgtatatat atatttatat
     4441 atagtcgatg tgcaaaagcc tacattattt ttcttttgat ttttattttt ttgttcaagt
     4501 aggttcatgc cgacagtaaa tccttatgcc ctacctaaat ataaacaccc aaatatatat
     4561 atatatatat atatatataa ataataaaaa cggaaaggaa atagaataat aataataata
     4621 ctacacgttt acgtatgtaa ctcattctat tattcagact ttgatattaa tagaattatt
     4681 actactaggt ttcctcactt caaaatctat aaatataccg tcatgctcta atcgatgctc
     4741 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4801 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4861 ataggctccg cccccgtgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4921 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4981 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5041 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5101 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5161 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5221 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5281 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5341 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5401 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5461 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5521 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5581 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5641 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5701 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5761 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5821 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5881 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5941 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6001 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6061 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6121 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6181 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6241 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6301 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6361 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6421 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6481 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6541 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6601 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     6661 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6721 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6781 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     6841 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6901 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6961 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     7021 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
     7081 tccaccgcgg tggcggccgc tctagaacta gtggatccat tgagtcatca gcctctctga
     7141 aaaaacgatt ctgaaataac cggaacttgt ttagattcct ttttgtaggt taatcttatc
     7201 ctgtacttgc tgatattgtg gttattagca gagatttttt tcaaaacatc atctaaagta
     7261 ggctttttgg ataagtcaat ttcagtgtcc cttaaccctt tagagcggct ttttatggtg
     7321 ataggcattt tcaaattaaa ttcaaaatat gtatctctct caaatagctc aattggtttc
     7381 tagatagcta aatagaatat aatcttactg tcctccgttc tgtaaaattc acgctcttag
     7441 tcccttttca taattcctta actttttgcg tacaaaatga tatgtttatt atatttttct
     7501 tttttttttt tcaaattttt tctttttctt gaaaaatttt tcaaattgga aagctcatct
     7561 ctcttgaatg tataatactt tcttcctcta actttcaaaa agttttacat agccaagaag
     7621 ttttccttac atcggtatac tactgttata taagttattc ttcgagaaac aattagatat
     7681 cattcatcgg ataaatctaa gttgcccatt gctttcaata actccgatca aattaactca
     7741 aatcaactaa aacagtaggg cccctcgaga tggtttctaa aggtgaagaa gataatatgg
     7801 ctattattaa agaatttatg agatttaaag ttcatatgga aggttctgtt aatggtcatg
     7861 aatttgaaat tgaaggtgaa ggtgaaggta gaccatatga aggtactcaa actgctaaat
     7921 tgaaagttac taaaggtggt ccattgccat ttgcttggga tattttgtct ccacaattta
     7981 tgtatggttc taaagcttat gttaaacatc cagctgatat tccagattat ttgaaattgt
     8041 cttttccaga aggttttaaa tgggaaagag ttatgaattt tgaagatggt ggtgttgtta
     8101 ctgttactca agattcttct ttgcaagatg gtgaatttat ttataaagtt aaattgagag
     8161 gtactaattt tccatctgat ggtccagtta tgcaaaaaaa aactatgggt tgggaagctt
     8221 cttctgaaag aatgtatcca gaagatggtg ctttgaaagg tgaaattaaa caaagattga
     8281 aattgaaaga tggtggtcat tatgatgctg aagttaaaac tacttataaa gctaaaaaac
     8341 cagttcaatt gccaggtgct tataatgtta atattaaatt ggatattact tctcataatg
     8401 aagattatac tatgttgaac atatgaaaga gctgaagtag acattctact ggtgtatgga
     8461 tgattgtata aaggtggtgg tgctcatcca gcatgggtat attcatattg gttgcactcc
     8521 ttatcttggt tttaggatgg ttctacagat aaatgagaga tcatatggtt ttgcatgaat
     8581 atgttaatgc tgctggtatt actggtggtg gttctggtgg tggttcttaa gttgcccttt
     8641 ccttctttct taccaacttt aatttctaag gcattgacat tgaaattgac attgtcctct
     8701 aatgtttata gagcaccatt atttgttttc gttttatata ttattatata atggcacaga
     8761 ataatttata attctttttt tttaagtcat tgaacagtcg cactaaccaa aagattaaac
     8821 accaccagag agtagttttt ccagcctggc aatctcttct aaagaagtag cttctgctaa
     8881 ctgctttttc aattcatttc ttctctctac tgtcatctta ctcaccacca aattcatgat
     8941 ctccatcgtt ttatctctgg atccttgt
//
