LOCUS       sequence-108805-        6549 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 6549)
  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 6549)
  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-1
FEATURES             Location/Qualifiers
     source          1..6549
                     /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             1921..2508
                     /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    2619..2700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2824..2905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3073..3528)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3210..3231)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3422..3441
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3555..3659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3660..3728,3729..4520)
                     /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(3878..3897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4691..5279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5180..5199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5433..5450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        5607..5910
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5912..6187
                     /label=chicken beta-actin promoter
     primer_bind     6443..6462
                     /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 cgccagccac gatggcggca agcaagcgct cgaaacggtg
      541 cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct
      601 atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
      661 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacggtggc
      721 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
      781 ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa
      841 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg
      901 gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg
      961 ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac
     1021 gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
     1081 catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg
     1141 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac
     1201 caagtggtgg ctatcgccag caacaatggc ggcaagcaag cgctcgaaac ggtgcagcgg
     1261 ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc
     1321 agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
     1381 caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag
     1441 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc
     1501 ccggaccaag tggtggctat cgccagcaac aatggcggca agcaagcgct cgaaacggtg
     1561 cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct
     1621 atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
     1681 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacaatggc
     1741 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
     1801 ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa
     1861 agcattgtgg cccagctgag ccggcctgat ccggcgttgg ccgcgttgac caacggatcc
     1921 cagctagtga aatctgaatt ggaagagaag aaatctgaac ttagacataa attgaaatat
     1981 gtgccacatg aatatattga attgattgaa atcgcaagaa attcaactca ggatagaatc
     2041 cttgaaatga aggtgatgga gttctttatg aaggtttatg gttatcgtgg taaacatttg
     2101 ggtggatcaa ggaaaccaga cggagcaatt tatactgtcg gatctcctat tgattacggt
     2161 gtgatcgttg atactaaggc atattcagga ggttataatc ttccaattgg tcaagcagat
     2221 gaaatgcaaa gatatgtcga agagaatcaa acaagaaaca agcatatcaa ccctaatgaa
     2281 tggtggaaag tctatccatc ttcagtaaca gaatttaagt tcttgtttgt gagtggtcat
     2341 ttcaaaggaa actacaaagc tcagcttaca agattgaatc atatcactaa ttgtaatgga
     2401 gctgttctta gtgtagaaga gcttttgatt ggtggagaaa tgattaaagc tggtacattg
     2461 acacttgagg aagtgagaag gaaatttaat aacggtgaga taaactttta ataggagctc
     2521 gcccggggat ctaattcaat tagagactaa ttcaattaga gctaattcaa ttaggatcca
     2581 gaattcattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2641 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2701 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2761 taaagcaagt aaaacctcta caaatgtggt cattgatgag tttggacaaa ccacaactag
     2821 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2881 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2941 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtcattc
     3001 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3061 aatggcgaat ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3121 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3181 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3241 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3301 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3361 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3421 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3481 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3541 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3601 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3661 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3721 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3781 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3841 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3901 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3961 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4021 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4081 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4141 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4201 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4261 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4321 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4381 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4441 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4501 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4561 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4621 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4681 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4741 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4801 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4861 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4921 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4981 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5041 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5101 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5161 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5221 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5281 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5341 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5401 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5461 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgag
     5521 atcttaaggg attactagtt taaacctagt tattaatagt aatcaattac ggggtcatta
     5581 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     5641 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     5701 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     5761 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     5821 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     5881 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     5941 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     6001 tgtgcagcga tgggggcggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga
     6061 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
     6121 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
     6181 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
     6241 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
     6301 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
     6361 gcgtgaaagc cttgaggggc tccgggagct agagcctctg ctaaccatgt tcatgccttc
     6421 ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc
     6481 aaagaatttt aagctaggat ctaactcgag aaagatattg tatatatcgt aacaatagga
     6541 ggttcaaca
//
