LOCUS       sequence-128833-        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N Trans V2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  AUTHORS   Slomovic S, Collins JJ
  TITLE     DNA sense-and-respond protein modules for mammalian cells.
  JOURNAL   Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585.
  PUBMED    26389572
REFERENCE   2  (bases 1 to 7793)
  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: "Nat 
            Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585."
COMMENT     Sequence Label: pVITRO1-SS-288
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(74..94)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        256..1562
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          583..1530
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     polyA_signal    1607..1728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1618..1637)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1672..1691
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1916..2504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2405..2424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2584..2887
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3002..4307
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3344..4275
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             4323..4346
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4347..4367
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5836..6280
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        6313..6359
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6351..6369
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6379..7173
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6379..7173
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6433..6452)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7043..7062
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7220..7792
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 cctgcagggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
       61 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      241 ccactagtgg agccgagagt aattcataca aaaggaggga tcgccttcgc aaggggagag
      301 cccagggacc gtccctaaat tctcacagac ccaaatccct gtagccgccc cacgacagcg
      361 cgaggagcat gcgctcaggg ctgagcgcgg ggagagcaga gcacacaagc tcatagaccc
      421 tggtcgtggg ggggaggacc ggggagctgg cgcggggcaa actgggaaag cggtgtcgtg
      481 tgctggctcc gccctcttcc cgagggtggg ggagaacggt atataagtgc ggcagtcgcc
      541 ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc aggtgagggg cgggtgtggc
      601 ttccgcgggc cgccgagctg gaggtcctgc tccgagcggg ccgggccccg ctgtcgtcgg
      661 cggggattag ctgcgagcat tcccgcttcg agttgcgggc ggcgcgggag gcagagtgcg
      721 aggcctagcg gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc
      781 cgcgccgccg ccgcgtgcta ctccggccgc actctggtct tttttttttt tgttgttgtt
      841 gccctgctgc cttcgattgc cgttcagcaa taggggctaa caaagggagg gtgcggggct
      901 tgctcgcccg gagcccggag aggtcatggt tggggaggaa tggagggaca ggagtggcgg
      961 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgg
     1021 ggtttttccc gaagcaacca ggctggggtt agcgtgccga ggccatgtgg ccccagcacc
     1081 cggcacgatc tggcttggcg gcgccgcgtt gccctgcctc cctaactagg gtgaggccat
     1141 cccgtccggc accagttgcg tgcgtggaaa gatggccgct cccgggccct gttgcaagga
     1201 gctcaaaatg gaggacgcgg cagcccggtg gagcgggcgg gtgagtcacc cacacaaagg
     1261 aagagggcct ggtccctcac cggctgctgc ttcctgtgac cccgtggtcc tatcggccgc
     1321 aatagtcacc tcgggctttt gagcacggct agtcgcggcg gggggagggg atgtaatggc
     1381 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
     1441 catttttgga atttgtcccc ttgagttttg agcggagcta attctcgggc ttcttagcgg
     1501 ttcaaaggta tcttttaaac ccttttttag gtgttgtgaa aaccaccgct aattcaaagc
     1561 aaccggtgat atcaaggatc ctgtacagct agctggccag acatgataag atacattgat
     1621 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1681 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1741 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1801 aacctctaca aatgtggtat ggaaatgtta attaactagc catgaccaaa atcccttaac
     1861 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1921 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1981 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2041 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2101 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2161 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2221 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2281 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2341 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2401 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2461 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2521 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttctta attaacctgc
     2581 aggcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     2641 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     2701 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     2761 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     2821 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     2881 taccatgatg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     2941 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttgac tagtggagcc
     3001 gagagtaatt catacaaaag gagggatcgc cttcgcaagg ggagagccca gggaccgtcc
     3061 ctaaattctc acagacccaa atccctgtag ccgccccacg acagcgcgag gagcatgcgc
     3121 ccagggctga gcgcgggtag atcagagcac acaagctcac agtccccggc ggtgggggga
     3181 ggggcgcgct gagcgggggc cagggagctg gcgcggggca aactgggaaa gtggtgtcgt
     3241 gtgctggctc cgccctcttc ccgagggtgg gggagaacgg tatataagtg cggtagtcgc
     3301 cttggacgtt ctttttcgca acgggtttgc cgtcagaacg caggtgagtg gcgggtgtgg
     3361 cttccgcggg ccccggagct ggagccctgc tctgagcggg ccgggctgat atgcgagtgt
     3421 cgtccgcagg gtttagctgt gagcattccc acttcgagtg gcgggcggtg cgggggtgag
     3481 agtgcgaggc ctagcggcaa ccccgtagcc tcgcctcgtg tccggcttga ggcctagcgt
     3541 ggtgtccgcc gccgcgtgcc actccggccg cactatgcgt tttttgtcct tgctgccctc
     3601 gattgccttc cagcagcatg ggctaacaaa gggagggtgt ggggctcact cttaaggagc
     3661 ccatgaagct tacgttggat aggaatggaa gggcaggagg ggcgactggg gcccgcccgc
     3721 cttcggagca catgtccgac gccacctgga tggggcgagg cctgtggctt tccgaagcaa
     3781 tcgggcgtga gtttagccta cctgggccat gtggccctag cactgggcac ggtctggcct
     3841 ggcggtgccg cgttcccttg cctcccaaca agggtgaggc cgtcccgccc ggcaccagtt
     3901 gcttgcgcgg aaagatggcc gctcccgggg ccctgttgca aggagctcaa aatggaggac
     3961 gcggcagccc ggtggagcgg gcgggtgagt cacccacaca aaggaagagg gccttgcccc
     4021 tcgccggccg ctgcttcctg tgaccccgtg gtctatcggc cgcatagtca cctcgggctt
     4081 ctcttgagca ccgctcgtcg cggcgggggg aggggatcta atggcgttgg agtttgttca
     4141 catttggtgg gtggagacta gtcaggccag cctggcgctg gaagtcattc ttggaatttg
     4201 cccctttgag tttggagcga ggctaattct caagcctctt agcggttcaa aggtattttc
     4261 taaacccgtt tccaggtgtt gtgaaagcca ccgctaattc aaagcaatcc ggagtataca
     4321 tggactacaa ggacgacgat gacaagccca agaaaaagcg caaggtctct agacccgggg
     4381 agcgcccctt ccagtgtcgc atttgcatgc ggaacttttc ggataaaact aaattgagag
     4441 ttcatacccg tactcatacc ggtgaaaaac cgtttcagtg tcggatctgt atgcgaaatt
     4501 tctccgttag acataatttg actagacatc tacgtacgca caccggcgag aagccattcc
     4561 aatgccgaat atgcatgcgc aacttcagtc aatctacttc tttgcaaaga cacctaaaaa
     4621 cccacctgag aggattcggc ggagtgctgg agaagggctg ctttgccaag ggtaccaatg
     4681 ttttaatggc ggatgggtct attgaatgta ttgaaaacat tgaggttggt aataaggtca
     4741 tgggtaaaga tggcagacct cgtgaggtaa ttaaattgcc cagaggaaga gaaactatgt
     4801 acagcgtcgt gcagaaaagt cagcacagag cccacaaaag tgactcaagt cgtgaagtgc
     4861 cagaattact caagtttacg tgtaatgcga cccatgagtt ggttgttaga acacctcgta
     4921 gtgtccgccg tttgtctcgt accattaagg gtgtcgaata ttttgaagtt attacttttg
     4981 agatgggcca aaagaaagcc cccgacggta gaattgttga gcttgtcaag gaagtttcaa
     5041 agagctaccc aatatctgag gggcctgaga gagccaacga attagtagaa tcctatagaa
     5101 aggcttcaaa taaagcttat tttgagtgga ctattgaggc cagagatctt tctctgttgg
     5161 gttgccacgt tcgcaaagcc acttatcaga catacgcccc aatcggcggg ggttctgggg
     5221 gaggatcagg tggtgggtcc gggggaggaa gcgggggtgg ctctgggggt ggatcaacac
     5281 gtcccgggga gcgccccttc cagtgcagga tctgcatgag gaattttagc attccaaacc
     5341 accttgcccg gcacacacgg actcacacag gggagaagcc cttccagtgt agaatttgca
     5401 tgcggaactt cagccagtca gcccacttga aaagacacct gcggacccat actggcgaaa
     5461 agccctttca gtgccggatc tgcatgcgca acttctccca ggacgtttcc cttgtcaggc
     5521 acctgaagac acatcttcgg cagaaagatg gagagagacc attccagtgt cggatttgta
     5581 tgaggaactt tagttctgca caggctttgg ctaggcatac ccggacccac acaggtgaga
     5641 agccattcca gtgtcgcatc tgcatgcgga atttctccca gggaggcaac ctcactaggc
     5701 acctgcggac tcatacaggg gagaaacctt ttcagtgtag gatctgcatg cgcaattttt
     5761 cacagcatcc aaatttgacg cgtcacctga aaacccacct gagaggatcc taaaagcttg
     5821 atgggggatc ctaggagcag gtttccccaa tgacacaaaa cgtgcaactt gaaactccgc
     5881 ctggtctttc caggtctaga ggggtaacac tttgtactgc gtttggctcc acgctcgatc
     5941 cactggcgag tgttagtaac agcactgttg cttcgtagcg gagcatgacg gccgtgggaa
     6001 ctcctccttg gtaacaagga cccacggggc caaaagccac gcccacacgg gcccgtcatg
     6061 tgtgcaaccc cagcacggcg actttactgc gaaacccact ttaaagtgac attgaaactg
     6121 gtacccacac actggtgaca ggctaaggat gcccttcagg taccccgagg taacacgcga
     6181 cactcgggat ctgagaaggg gactggggct tctataaaag cgctcggttt aaaaagcttc
     6241 tatgcctgaa taggtgaccg gaggtcggca cctttccttt gcaattactg accctatgaa
     6301 tacactgact gtttgacaat taatcatcgg catagtatat cggcatagta taatacgact
     6361 cactatagga gggccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6421 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6481 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6541 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     6601 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6661 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6721 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6781 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6841 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6901 aaggcgagca tgcccgacgg cgaggatctc gtcgtgacac atggcgatgc ctgcttgccg
     6961 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     7021 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     7081 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     7141 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     7201 accaagcgaa ttcgctagga ttatccctaa tacctgccac cccactctta atcagtggtg
     7261 gaagaacggt ctcagaactg tttgtttcaa ttggccattt aagtttagta gtaaaagact
     7321 ggttaatgat aacaatgcat cgtaaaacct tcagaaggaa aggagaatgt tttgtggacc
     7381 actttggttt tcttttttgc gtgtggcagt tttaagttat tagtttttaa aatcagtact
     7441 ttttaatgga aacaacttga ccaaaaattt gtcacagaat tttgagaccc attaaaaaag
     7501 ttaaatgaga aacctgtgtg ttcctttggt caacaccgag acatttaggt gaaagacatc
     7561 taattctggt tttacgaatc tggaaacttc ttgaaaatgt aattcttgag ttaacacttc
     7621 tgggtggaga atagggttgt tttcccccca cataattgga aggggaagga atatcattta
     7681 aagctatggg agggttgctt tgattacaac actggagaga aatgcagcat gttgctgatt
     7741 gcctgtcact aaaacaggcc aaaaactgag tccttgggtt gcatagaaag ctg
//
