LOCUS       sequence-48254-e        5701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  destination vector for the Golden Gate TALEN kit, directs expression
            of TALENs from a truncated CAGs promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  AUTHORS   Carlson DF, Tan W, Lillico SG, Stverakova D, Proudfoot C, Christian 
            M, Voytas DF, Long CR, Whitelaw CB, Fahrenkrug SC
  TITLE     Efficient TALEN-mediated gene knockout in livestock.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Oct 1.
  PUBMED    23027955
REFERENCE   2  (bases 1 to 5701)
  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: "Proc Natl 
            Acad Sci U S A. 2012 Oct 1."
COMMENT     Sequence Label: pC-GoldyTALEN
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        104..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..684
                     /label=chicken beta-actin promoter
     primer_bind     940..959
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1062..1082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1083..1109
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             complement(1413..1820)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     1670..1692
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1684..1701
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1685..1701
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1711..1730
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1711..1729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1777..1797)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1777..1795)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1816..1832)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1816..1832)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1829..1851)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1840..1856
                     /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(1864..1870,1871..1888,1889..1894))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1909..1930
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2119..2706
                     /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    2817..2898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3022..3103
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3271..3726)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3408..3429)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3620..3639
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3753..3857
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3858..3926,3927..4718)
                     /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(4076..4095)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4889..5477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5378..5397
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5631..5648
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcattaatgc agctgagatc ttaagggatt actagtttaa acctagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
      421 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
      481 ttatttttta attattttgt gcagcgatgg gggcgggggg ggggggggcg cgcgccaggc
      541 ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat
      601 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
      661 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
      721 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      781 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
      841 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagctaga gcctctgcta
      901 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
      961 tgtctcatca ttttggcaaa gaattttaag ctaggatcta actcgagaaa gatattgtat
     1021 atatcgtaac aataggaggt tcaacaatgg cttcctcccc tccaaagaaa aagagaaagg
     1081 ttagttggaa ggacgcaagt ggttggtcta gagtggatct acgcacgctc ggctacagtc
     1141 agcagcagca agagaagatc aaaccgaagg tgcgttcgac agtggcgcag caccacgagg
     1201 cactggtggg ccatgggttt acacacgcgc acatcgttgc gctcagccaa cacccggcag
     1261 cgttagggac cgtcgctgtc acgtatcagc acataatcac ggcgttgcca gaggcgacac
     1321 acgaagacat cgttggcgtc ggcaaacagt ggtccggcgc acgcgccctg gaggccttgc
     1381 tcacggatgc gggggagttg agaggtccgc cgttacagtt ggacacaggc caacttgtga
     1441 agattgcaaa acgtggcggc gtgaccgcaa tggaggcagt gcatgcatcg cgcaatgcac
     1501 tgacgggtgc ccccctggag acgggcgccg ctacagggcg cgtcccattc gccattcagg
     1561 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     1621 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     1681 cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt
     1741 accgggcccc ccctcgaggt cctccagctt ttgttccctt tagtgagggt taattgcgcg
     1801 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1861 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1921 actcacatta attgcgttgc gctcactgcc cgctttccac cggtcgtctc caacgaccac
     1981 ctcgtcgcct tggcctgcct cggcggacgt cctgccatgg atgcagtgaa aaagggattg
     2041 ccgcacgcgc cggaattgat cagaagagtc aatcgccgta ttggcgaacg cacgtcccat
     2101 cgcgttgcct ctagatccca gctagtgaaa tctgaattgg aagagaagaa atctgaactt
     2161 agacataaat tgaaatatgt gccacatgaa tatattgaat tgattgaaat cgcaagaaat
     2221 tcaactcagg atagaatcct tgaaatgaag gtgatggagt tctttatgaa ggtttatggt
     2281 tatcgtggta aacatttggg tggatcaagg aaaccagacg gagcaattta tactgtcgga
     2341 tctcctattg attacggtgt gatcgttgat actaaggcat attcaggagg ttataatctt
     2401 ccaattggtc aagcagatga aatgcaaaga tatgtcgaag agaatcaaac aagaaacaag
     2461 catatcaacc ctaatgaatg gtggaaagtc tatccatctt cagtaacaga atttaagttc
     2521 ttgtttgtga gtggtcattt caaaggaaac tacaaagctc agcttacaag attgaatcat
     2581 atcactaatt gtaatggagc tgttcttagt gtagaagagc ttttgattgg tggagaaatg
     2641 attaaagctg gtacattgac acttgaggaa gtgagaagga aatttaataa cggtgagata
     2701 aacttttaat aggagctcgc ccggggatct aattcaatta gagactaatt caattagagc
     2761 taattcaatt aggatccaga attcattgat gagtttggac aaaccacaac tagaatgcag
     2821 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2881 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2941 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtca ttgatgagtt
     3001 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3061 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3121 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3181 ctacaaatgt ggtcattcag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3241 caacagttgc gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat
     3301 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3361 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3421 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3481 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3541 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3601 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3661 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3721 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3781 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3841 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3901 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3961 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4021 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4081 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4141 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4201 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4261 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4321 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4381 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4441 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4501 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4561 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4621 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4681 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4741 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4801 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4861 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4921 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4981 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5041 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5101 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5161 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5221 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5281 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5341 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5401 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5461 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5521 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5581 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5641 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5701 t
//
