LOCUS       sequence-350825-        6146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tobacco etch virus protease (TEV).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6146)
  AUTHORS   To TL, Piggott BJ, Makhijani K, Yu D, Jan YN, Shu X
  TITLE     Rationally designed fluorogenic protease reporter visualizes 
            spatiotemporal dynamics of apoptosis in vivo.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3338-43. doi: 
            10.1073/pnas.1502857112. Epub 2015 Mar 2.
  PUBMED    25733847
REFERENCE   2  (bases 1 to 6146)
  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"; doi: 
            "10.1073/pnas.1502857112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-03-17- 17"; volume: "112"; issue: "11"; pages: "3338-43"
COMMENT     Sequence Label: pcDNA3.1 TEV (full-length)
FEATURES             Location/Qualifiers
     source          1..6146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(112..132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        117..446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      297..432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     359..378
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             513..1307
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(567..586)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1177..1196
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1483..1604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1520..1539)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1574..1593
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1653..1669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1653..1669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1666..1688)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1677..1693
                     /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(1701..1707,1708..1725,1726..1731))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1746..1767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1884..1901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2055..2643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2135..2154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2814..3605,3606..3674))
                     /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     3437..3456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3675..3779)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3854..3873)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4045..4424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4425..4628
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4579..4599
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4673..4692
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4673..4691
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4718..4727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5460..5489
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5505..5522
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5545..5562)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5551..5775
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5821..6146,1..103)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5908..5927)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6118..6139
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
       61 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
      121 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      181 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
      241 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
      301 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
      361 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
      421 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
      481 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
      541 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
      601 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
      661 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
      721 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
      781 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
      841 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
      901 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
      961 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     1021 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     1081 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     1141 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     1201 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     1261 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     1321 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     1381 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     1441 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     1501 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     1561 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     1621 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1681 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1741 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1801 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1861 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1921 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1981 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2041 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2101 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2161 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2221 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2281 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2341 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2401 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2461 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2521 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2581 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2641 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2701 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2761 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2821 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2881 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2941 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3001 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3061 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3121 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3181 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3241 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3301 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3361 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3421 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3481 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3541 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3601 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3661 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3721 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3781 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     3841 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     3901 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     3961 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     4021 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
     4081 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     4141 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     4201 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     4261 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     4321 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     4381 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     4441 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4501 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4561 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     4621 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
     4681 ctcactatag ggagacccaa gctggctagt taagcttgcc accatgggcg agagcctttt
     4741 caagggcccg agggactaca acccgatctc cagcaccatc tgtcacctga ccaacgagag
     4801 cgacggtcac accactagtc tgtacggcat cggcttcggc cccttcatca tcaccaacaa
     4861 gcatctgttc aggaggaata acggcacact gctggtgcaa agcctgcacg gcgtgttcaa
     4921 agtgaagaac acaaccaccc tgcaacagca cctgatcgac ggcagggaca tgattatcat
     4981 caggatgccc aaggacttcc ccccctttcc ccagaaactg aagttcaggg agccacaaag
     5041 ggaggagcga atctgcctgg tgaccaccaa cttccagacc aagtccatga gcagcatggt
     5101 ctctgatacc agctgcacct tccccagcag cgacggcatc ttctggaagc actggattca
     5161 gacgaaggat ggccaatgcg gcagcccatt ggtgagcact agggacggct tcatcgtggg
     5221 catccacagc gccagcaatt ttaccaatac caacaactac ttcacgagcg tgccgaaaaa
     5281 cttcatggag ctgttgacca atcaagaggc gcagcagtgg gtgagcggct ggaggctgaa
     5341 cgccgacagc gttctttggg gcggacataa ggtgttcatg gtcaagcccg aggaaccctt
     5401 ccagcccgtt aaggaagcca ctcagctttg ataactcgag tctagagggc ccgcggttcg
     5461 aacaaaaact catctcagaa gaggatctga atatgcatac cggtcatcat caccatcacc
     5521 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     5581 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     5641 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     5701 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     5761 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     5821 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5881 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5941 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6001 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6061 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6121 gactcttgtt ccaaactgga acaaca
//
