LOCUS       sequence-121038-        7336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C Trans V3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7336)
  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 7336)
  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-259
FEATURES             Location/Qualifiers
     source          1..7336
                     /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"
     CDS             1585..1608
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1609..1629
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2343..2354
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2467..2616
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     polyA_signal    2648..2769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2659..2678)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2713..2732
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2957..3545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3446..3465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3625..3928
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4043..5348
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    4385..5316
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     misc_feature    5379..5823
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        5856..5902
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5894..5912
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5922..6716
                     /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             5922..6716
                     /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(5976..5995)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6586..6605
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6763..7335
                     /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 catggactac aaagacgatg acgacaagcc taagaagaaa
     1621 cggaaggtgt ccagacctgg ggagaggcca ttccagtgtc ggatctgtat gagaaatttt
     1681 agcactatgg ctgtactgag gagacataca agaacccata caggcgagaa gccctttcag
     1741 tgcaggatct gcatgcgaaa cttttctcgc agagaagtgc ttgaaaacca cctgcggact
     1801 cataccggcg aaaaaccatt tcagtgtcga atttgcatgc gcaactttag ccagacagtg
     1861 aacttggata gacaccttaa gactcatctg aggcagaagg acggggagag acctttccag
     1921 tgcagaatct gcatgagaaa cttttctaag aaagatcatc tgcaccgcca tacccgcacc
     1981 cacaccggcg aaaaaccctt ccagtgccgc atctgtatgc gaaattttag ccagaggcca
     2041 catcttacaa atcaccttag aacacacact ggagaaaaac catttcagtg tcggatctgc
     2101 atgcgcaact tctccgtggg cgccagcctc aaacgccacc tgaaaaccca cctgagggga
     2161 tctgggggcg gatcaggtgg aggctcagga ggtgggtccg gaggtggatc tggtggcggg
     2221 tccggaggcg gatccgtttt gcttaacgtt ctttcgaagt gtgccggctc taaaaaattc
     2281 aggcctgctc ccgccgctgc ttttgcacgt gagtgccgcg gattttattt cgagttacaa
     2341 gaattgaagg aagacgatta ttatgggatt actttatctg atgattctga tcatcagttt
     2401 ttgcttgcca accaggttgt cgtccataat tgcgacagcg tgctgtgggg cagcggaggg
     2461 cgcgccgacg cgctggacga tttcgatctc gacatgctgg gttctgatgc cctcgatgac
     2521 tttgacctgg atatgttggg aagcgacgca ttggatgact ttgatctgga catgctcggc
     2581 tccgatgctc tggacgattt cgatctcgat atgttaatta attgttaagc tagctggcca
     2641 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2701 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2761 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2821 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggaaatgtt aattaactag
     2881 ccatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2941 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3001 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3061 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3121 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3181 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3241 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3301 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3361 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3421 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3481 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3541 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3601 acatgttctt aattaacctg caggcgttac ataacttacg gtaaatggcc cgcctggctg
     3661 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     3721 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     3781 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     3841 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     3901 ctacgtatta gtcatcgcta ttaccatgat gatgcggttt tggcagtaca tcaatgggcg
     3961 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     4021 tttgttttga ctagtggagc cgagagtaat tcatacaaaa ggagggatcg ccttcgcaag
     4081 gggagagccc agggaccgtc cctaaattct cacagaccca aatccctgta gccgccccac
     4141 gacagcgcga ggagcatgcg cccagggctg agcgcgggta gatcagagca cacaagctca
     4201 cagtccccgg cggtgggggg aggggcgcgc tgagcggggg ccagggagct ggcgcggggc
     4261 aaactgggaa agtggtgtcg tgtgctggct ccgccctctt cccgagggtg ggggagaacg
     4321 gtatataagt gcggtagtcg ccttggacgt tctttttcgc aacgggtttg ccgtcagaac
     4381 gcaggtgagt ggcgggtgtg gcttccgcgg gccccggagc tggagccctg ctctgagcgg
     4441 gccgggctga tatgcgagtg tcgtccgcag ggtttagctg tgagcattcc cacttcgagt
     4501 ggcgggcggt gcgggggtga gagtgcgagg cctagcggca accccgtagc ctcgcctcgt
     4561 gtccggcttg aggcctagcg tggtgtccgc cgccgcgtgc cactccggcc gcactatgcg
     4621 ttttttgtcc ttgctgccct cgattgcctt ccagcagcat gggctaacaa agggagggtg
     4681 tggggctcac tcttaaggag cccatgaagc ttacgttgga taggaatgga agggcaggag
     4741 gggcgactgg ggcccgcccg ccttcggagc acatgtccga cgccacctgg atggggcgag
     4801 gcctgtggct ttccgaagca atcgggcgtg agtttagcct acctgggcca tgtggcccta
     4861 gcactgggca cggtctggcc tggcggtgcc gcgttccctt gcctcccaac aagggtgagg
     4921 ccgtcccgcc cggcaccagt tgcttgcgcg gaaagatggc cgctcccggg gccctgttgc
     4981 aaggagctca aaatggagga cgcggcagcc cggtggagcg ggcgggtgag tcacccacac
     5041 aaaggaagag ggccttgccc ctcgccggcc gctgcttcct gtgaccccgt ggtctatcgg
     5101 ccgcatagtc acctcgggct tctcttgagc accgctcgtc gcggcggggg gaggggatct
     5161 aatggcgttg gagtttgttc acatttggtg ggtggagact agtcaggcca gcctggcgct
     5221 ggaagtcatt cttggaattt gcccctttga gtttggagcg aggctaattc tcaagcctct
     5281 tagcggttca aaggtatttt ctaaacccgt ttccaggtgt tgtgaaagcc accgctaatt
     5341 caaagcaatc cggagtatac ggatcccgta cgcctaggag caggtttccc caatgacaca
     5401 aaacgtgcaa cttgaaactc cgcctggtct ttccaggtct agaggggtaa cactttgtac
     5461 tgcgtttggc tccacgctcg atccactggc gagtgttagt aacagcactg ttgcttcgta
     5521 gcggagcatg acggccgtgg gaactcctcc ttggtaacaa ggacccacgg ggccaaaagc
     5581 cacgcccaca cgggcccgtc atgtgtgcaa ccccagcacg gcgactttac tgcgaaaccc
     5641 actttaaagt gacattgaaa ctggtaccca cacactggtg acaggctaag gatgcccttc
     5701 aggtaccccg aggtaacacg cgacactcgg gatctgagaa ggggactggg gcttctataa
     5761 aagcgctcgg tttaaaaagc ttctatgcct gaataggtga ccggaggtcg gcacctttcc
     5821 tttgcaatta ctgaccctat gaatacactg actgtttgac aattaatcat cggcatagta
     5881 tatcggcata gtataatacg actcactata ggagggccac catgattgaa caagatggat
     5941 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     6001 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     6061 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     6121 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6181 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6241 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6301 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6361 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6421 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     6481 cacatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6541 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6601 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6661 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6721 gactctgggg ttcgaaatga ccgaccaagc gaattcgcta ggattatccc taatacctgc
     6781 caccccactc ttaatcagtg gtggaagaac ggtctcagaa ctgtttgttt caattggcca
     6841 tttaagttta gtagtaaaag actggttaat gataacaatg catcgtaaaa ccttcagaag
     6901 gaaaggagaa tgttttgtgg accactttgg ttttcttttt tgcgtgtggc agttttaagt
     6961 tattagtttt taaaatcagt actttttaat ggaaacaact tgaccaaaaa tttgtcacag
     7021 aattttgaga cccattaaaa aagttaaatg agaaacctgt gtgttccttt ggtcaacacc
     7081 gagacattta ggtgaaagac atctaattct ggttttacga atctggaaac ttcttgaaaa
     7141 tgtaattctt gagttaacac ttctgggtgg agaatagggt tgttttcccc ccacataatt
     7201 ggaaggggaa ggaatatcat ttaaagctat gggagggttg ctttgattac aacactggag
     7261 agaaatgcag catgttgctg attgcctgtc actaaaacag gccaaaaact gagtccttgg
     7321 gttgcataga aagctg
//
