LOCUS       sequence-108809-        7161 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEN mediated gene editing at human TUBA1B locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7161)
  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 7161)
  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: pGoldyTALEN-TUBA1B-TAL-2
FEATURES             Location/Qualifiers
     source          1..7161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             16..36
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             37..63
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             2533..3120
                     /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    3231..3312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3436..3517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3685..4140)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3822..3843)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4034..4053
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4167..4271
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4272..4340,4341..5132)
                     /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(4490..4509)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5303..5891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5792..5811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6045..6062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        6219..6522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6524..6799
                     /label=chicken beta-actin promoter
     primer_bind     7055..7074
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 atggcttcct cccctccaaa gaaaaagaga aaggttagtt ggaaggacgc aagtggttgg
       61 tctagagtgg atctacgcac gctcggctac agtcagcagc agcaagagaa gatcaaaccg
      121 aaggtgcgtt cgacagtggc gcagcaccac gaggcactgg tgggccatgg gtttacacac
      181 gcgcacatcg ttgcgctcag ccaacacccg gcagcgttag ggaccgtcgc tgtcacgtat
      241 cagcacataa tcacggcgtt gccagaggcg acacacgaag acatcgttgg cgtcggcaaa
      301 cagtggtccg gcgcacgcgc cctggaggcc ttgctcacgg atgcggggga gttgagaggt
      361 ccgccgttac agttggacac aggccaactt gtgaagattg caaaacgtgg cggcgtgacc
      421 gcaatggagg cagtgcatgc atcgcgcaat gcactgacgg gtgcccccct gaacctgacc
      481 ccggaccaag tggtggctat cgccagcaac aatggcggca agcaagcgct cgaaacggtg
      541 cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct
      601 atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
      661 ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc
      721 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
      781 ctgaccccgg accaagtggt ggctatcgcc agcaacattg gcggcaagca agcgctcgaa
      841 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
      901 gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg
      961 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac
     1021 ggtggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
     1081 catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg
     1141 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac
     1201 caagtggtgg ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg
     1261 ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc
     1321 agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
     1381 caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacaa tggcggcaag
     1441 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc
     1501 ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct cgaaacggtg
     1561 cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct
     1621 atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
     1681 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc
     1741 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
     1801 ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa
     1861 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg
     1921 gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg
     1981 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac
     2041 attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
     2101 catggcctga ccccggacca agtggtggct atcgccagca acaatggcgg caagcaagcg
     2161 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac
     2221 caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg
     2281 ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc
     2341 agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
     2401 caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag
     2461 caagcgctcg aaagcattgt ggcccagctg agccggcctg atccggcgtt ggccgcgttg
     2521 accaacggat cccagctagt gaaatctgaa ttggaagaga agaaatctga acttagacat
     2581 aaattgaaat atgtgccaca tgaatatatt gaattgattg aaatcgcaag aaattcaact
     2641 caggatagaa tccttgaaat gaaggtgatg gagttcttta tgaaggttta tggttatcgt
     2701 ggtaaacatt tgggtggatc aaggaaacca gacggagcaa tttatactgt cggatctcct
     2761 attgattacg gtgtgatcgt tgatactaag gcatattcag gaggttataa tcttccaatt
     2821 ggtcaagcag atgaaatgca aagatatgtc gaagagaatc aaacaagaaa caagcatatc
     2881 aaccctaatg aatggtggaa agtctatcca tcttcagtaa cagaatttaa gttcttgttt
     2941 gtgagtggtc atttcaaagg aaactacaaa gctcagctta caagattgaa tcatatcact
     3001 aattgtaatg gagctgttct tagtgtagaa gagcttttga ttggtggaga aatgattaaa
     3061 gctggtacat tgacacttga ggaagtgaga aggaaattta ataacggtga gataaacttt
     3121 taataggagc tcgcccgggg atctaattca attagagact aattcaatta gagctaattc
     3181 aattaggatc cagaattcat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3241 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3301 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3361 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtcattgatg agtttggaca
     3421 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3481 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3541 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3601 atgtggtcat tcagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3661 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
     3721 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3781 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3841 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3901 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3961 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4021 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4081 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4141 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4201 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4261 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4321 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4381 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4441 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4501 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4561 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4621 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4681 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4741 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4801 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4861 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4921 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4981 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5041 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5101 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5161 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5221 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5281 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5341 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5401 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5461 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5521 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5581 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5641 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5701 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5761 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5821 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5881 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5941 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6001 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6061 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6121 aatgcagctg agatcttaag ggattactag tttaaaccta gttattaata gtaatcaatt
     6181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     6241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     6421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     6481 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     6541 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     6601 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggcgcgcgc caggcggggc
     6661 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
     6721 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
     6781 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
     6841 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     6901 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
     6961 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ctagagcctc tgctaaccat
     7021 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
     7081 catcattttg gcaaagaatt ttaagctagg atctaactcg agaaagatat tgtatatatc
     7141 gtaacaatag gaggttcaac a
//
