LOCUS       sequence-101922-        5563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression TALEN vector with N152, C+18 truncations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5563)
  AUTHORS   Gan Z, Ding L, Burckhardt CJ, Lowery J, Zaritsky A, Sitterley K, 
            Mota A, Costigliola N, Starker CG, Voytas DF, Tytell J, Goldman RD, 
            Danuser G
  TITLE     Vimentin Intermediate Filaments Template Microtubule Networks to 
            Enhance Persistence in Cell Polarity and Directed Migration.
  JOURNAL   Cell Syst. 2016 Sep 21. pii: S2405-4712(16)30262-9. doi: 
            10.1016/j.cels.2016.08.007.
  PUBMED    27667364
REFERENCE   2  (bases 1 to 5563)
  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: "Cell Syst. 
            2016 Sep 21. pii: S2405-4712(16)30262-9. doi: 
            10.1016/j.cels.2016.08.007."
COMMENT     Sequence Label: pCGS652
FEATURES             Location/Qualifiers
     source          1..5563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        362..637
                     /label=chicken beta-actin promoter
     primer_bind     893..912
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1015..1035
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1036..1062
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             complement(1366..1773)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     1623..1645
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1637..1654
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1638..1654
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1664..1683
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1664..1682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1730..1750)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1730..1748)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1769..1785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1769..1785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1782..1804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1793..1809
                     /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(1817..1823,1824..1841,1842..1847))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1862..1883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1934..2521
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     polyA_signal    2632..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2837..2918
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3086..3541)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3223..3244)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3435..3454
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3568..3672
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3673..3741,3742..4533)
                     /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     complement(3891..3910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4704..5292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5193..5212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5446..5463
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
       61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      361 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
      421 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
      481 gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc
      541 ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg
      601 gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg
      661 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
      721 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt
      781 ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggagct
      841 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
      901 ctggttattg tgctgtctca tcattttggc aaagaatttt aagctaggat ctaactcgag
      961 aaagatattg tatatatcgt aacaatagga ggttcaacaa tggcttcctc ccctccaaag
     1021 aaaaagagaa aggttagttg gaaggacgca agtggttggt ctagagtgga tctacgcacg
     1081 ctcggctaca gtcagcagca gcaagagaag atcaaaccga aggtgcgttc gacagtggcg
     1141 cagcaccacg aggcactggt gggccatggg tttacacacg cgcacatcgt tgcgctcagc
     1201 caacacccgg cagcgttagg gaccgtcgct gtcacgtatc agcacataat cacggcgttg
     1261 ccagaggcga cacacgaaga catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc
     1321 ctggaggcct tgctcacgga tgcgggggag ttgagaggtc cgccgttaca gttggacaca
     1381 ggccaacttg tgaagattgc aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca
     1441 tcgcgcaatg cactgacggg tgcccccctg gagacgggcg ccgctacagg gcgcgtccca
     1501 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     1561 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     1621 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     1681 gggcgaattg ggtaccgggc cccccctcga ggtcctccag cttttgttcc ctttagtgag
     1741 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1801 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1861 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc caccggtcgt
     1921 ctccaacgga tcccagctag tgaaatctga attggaagag aagaaatctg aacttagaca
     1981 taaattgaaa tatgtgccac atgaatatat tgaattgatt gaaatcgcaa gaaattcaac
     2041 tcaggataga atccttgaaa tgaaggtgat ggagttcttt atgaaggttt atggttatcg
     2101 tggtaaacat ttgggtggat caaggaaacc agacggagca atttatactg tcggatctcc
     2161 tattgattac ggtgtgatcg ttgatactaa ggcatattca ggaggttata atcttccaat
     2221 tggtcaagca gatgaaatgc aaagatatgt cgaagagaat caaacaagaa acaagcatat
     2281 caaccctaat gaatggtgga aagtctatcc atcttcagta acagaattta agttcttgtt
     2341 tgtgagtggt catttcaaag gaaactacaa agctcagctt acaagattga atcatatcac
     2401 taattgtaat ggagctgttc ttagtgtaga agagcttttg attggtggag aaatgattaa
     2461 agctggtaca ttgacacttg aggaagtgag aaggaaattt aataacggtg agataaactt
     2521 ttaataggag ctcgcccggg gatctaattc aattagagac taattcaatt agagctaatt
     2581 caattaggat ccagaattca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2641 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2701 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2761 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtcattgat gagtttggac
     2821 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2881 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2941 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     3001 aatgtggtca ttcagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3061 gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta atattttgtt aaaattcgcg
     3121 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3181 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3241 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3301 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3361 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3421 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3481 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3541 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3601 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3661 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3721 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3781 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3841 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3901 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3961 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4021 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4081 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4141 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4201 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4261 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4321 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4381 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4441 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4501 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4561 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4621 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4681 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4741 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4801 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4861 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4921 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4981 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5041 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5101 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5161 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5221 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5281 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5341 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5401 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5461 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     5521 taatgcagct gagatcttaa gggattacta gtttaaacct agt
//
