LOCUS       sequence-437089-        7156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector of eGFP tagged NFATC2IP ORF with nuclear
            export signal (NES) at its N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7156)
  AUTHORS   Cho T, Hoeg L, Setiaputra D, Durocher D
  TITLE     NFATC2IP is a mediator of SUMO-dependent genome integrity.
  JOURNAL   Genes Dev. 2024 Apr 17;38(5-6):233-252. doi: 10.1101/gad.350914.123.
  PUBMED    38503515
REFERENCE   2  (bases 1 to 7156)
  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: "Genes 
            Dev."; date: "2024-04-17"; volume: "38(5-6):233-252. doi"; pages: " 
            10.1101/gad.350914.123"
COMMENT     Sequence Label: pcDNA5-FRT/T0-eGFP-NES-NFATC2IP
FEATURES             Location/Qualifiers
     source          1..7156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(990..992,993..995,996..1706)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(990..992,993..995,996..1706)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1035..1056)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1296..1315)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1643..1664
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3108..3125)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3114..3338
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3471..3490)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3622..3669
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3677..4699
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4829..4950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4866..4885)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4920..4939
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4999..5015)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4999..5015)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5012..5034)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5023..5039
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5047..5053,5054..5071,5072..5077))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5092..5113
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5230..5247)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5401..5989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5481..5500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6160..6951,6952..7020))
                     /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     6783..6802
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7021..7125)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacca tggtgagcaa gggcgaggag ctgttcaccg
     1021 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1081 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1141 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1201 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1261 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1321 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1381 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1441 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1501 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1561 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1621 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1681 ctctcggcat ggacgagctg tacaagggcg cgccaaatct cgtggacctc caaaagaagc
     1741 tggaggagct ggagctggac gagcagcagg gcgctggagc cggcgcgcca gcggagcctg
     1801 tggggaagcg gggccgctgg tccggaggta gcggtgccgg ccgagggggt cggggcggct
     1861 ggggcggtcg gggccggcgt cctcgggccc agcggtctcc atcccggggc acgctggacg
     1921 tagtgtctgt ggacttggtc accgacagcg atgaggaaat tctggaggtc gccaccgctc
     1981 gcggtgccgc ggacgaggtt gaggtggagc ccccggagcc cccggggccg gtcgcgtccc
     2041 gggataacag caacagtgac agcgaagggg aggacaggcg gcccgcagga cccccgcggg
     2101 agccggtcag gcggcggcgg cggctggtgc tggatccggg ggaggcgccg ctggttccgg
     2161 tgtactcggg gaaggttaaa agcagccttc gccttatccc agatgatcta tccctcctga
     2221 aactctaccc tccaggggat gaggaagagg cagagctggc agattcgagt ggtctctacc
     2281 atgagggctc cccatcacca ggctctccct ggaagacaaa gctgaggact aaggataaag
     2341 aagagaagaa aaagacagag tttctggatc tggacaactc tcctctgtcc ccaccttcac
     2401 caaggaccaa aagcagaacg catactcggg cactcaagaa gttaagtgag gtgaacaagc
     2461 gcctccagga tctccgttcc tgtctgagcc ccaagccacc tcagggtcaa gagcaacagg
     2521 gccaagagga tgaagtggtc ttggtggaag ggcccaccct cccagagacc ccccgactct
     2581 tcccactcaa aatccgttgc cgggctgacc tggtcagatt gcccctcagg atgtcggagc
     2641 ccctgcagag tgtggtggac cacatggcca cccaccttgg ggtgtcccca agcaggatcc
     2701 ttttgctttt tggagagaca gagctatcac ctactgccac tcccaggacc ctaaagctcg
     2761 gagtggctga catcattgac tgtgtggtac taacaagttc tccagaggcc acagagacgt
     2821 cccaacagct ccagctccgg gtgcagggaa aggagaaaca ccagacactg gaagtctcac
     2881 tgtctcgaga ttcccctcta aagaccctca tgtcccacta tgaggaggcc atgggactgt
     2941 cgggacggaa gctctccttc ttctttgatg ggacaaagct ttcaggcagg gagctgccag
     3001 ctgacctggg catggaatct ggggacctca ttgaggtctg gggctgagat atccatcaca
     3061 ctggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3121 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3181 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3241 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3301 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3361 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3421 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3481 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3541 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3601 gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat
     3661 aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3721 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     3781 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     3841 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     3901 agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     3961 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     4021 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     4081 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4141 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4201 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4261 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4321 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4381 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4441 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4501 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4561 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     4621 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4681 tccgagggca aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga
     4741 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4801 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4861 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4921 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4981 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5041 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5101 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5161 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5221 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5281 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5341 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5401 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5461 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5521 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5581 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5641 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5701 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5761 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5821 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5881 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5941 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6001 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6061 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6121 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6181 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6241 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6301 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6361 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6421 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6481 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6541 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6601 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6661 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6721 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6781 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6841 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6901 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6961 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7021 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7081 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7141 agtgccacct gacgtc
//
