LOCUS       Exported                4738 bp ds-DNA     linear   SYN 18-JUL-2021
DEFINITION  Destination vector for TALE-KRAB construction.
ACCESSION   .
VERSION     .
KEYWORDS    pTALE-STAR-KRAB-G
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4738)
  AUTHORS   Gogolok S, Garcia-Diaz C, Pollard SM
  TITLE     STAR: a simple TAL effector assembly reaction using isothermal 
            assembly.
  JOURNAL   Sci Rep. 2016 Sep 12;6:33209. doi: 10.1038/srep33209.
  PUBMED    27615025
REFERENCE   2  (bases 1 to 4738)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 18, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4738
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     39..61
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     123..142
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      179..188
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             188..253
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             260..280
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1063..1083
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1138..1332
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     primer_bind     complement(1355..1372)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1361..1585
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1632..1648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1632..1648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1645..1667)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1656..1672
                     /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(1680..1710)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1725..1746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1805..2000
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1851..1986
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1913..1932
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2006..2140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2208..2225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2379..2967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2459..2478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3138..3998)
                     /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     3761..3780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4591..4725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
       61 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
      121 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcc tcgaggccgc
      181 caccatggac tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga
      241 tgacgatgac aagatggccc ccaagaagaa gaggaaggtg ggcattcacc gcggggtacc
      301 tatggtggac ttgaggacac tcggttattc gcaacagcaa caggagaaaa tcaagcctaa
      361 ggtcaggagc accgtcgcgc aacaccacga ggcgcttgtg gggcatggct tcactcatgc
      421 gcatattgtc gcgctttcac agcaccctgc ggcgcttggg acggtggctg tcaaatacca
      481 agatatgatt gcggccctgc ccgaagccac gcacgaggca attgtagggg tcggtaaaca
      541 gtggtcggga gcgcgagcac ttgaggcgct gctgactgtg gcgggtgagc ttagggggcc
      601 tccgctccag ctcgacaccg ggcagctgct gaagatcgcg aagagagggg gagtaacagc
      661 ggtagaggca gtgcacgcct ggcgcaatgc gctcaccggg gcccccttga acctgcagga
      721 tacgtttatc agtgataaag tgtcaagcat gacaaagttg cagccgaata cagtggtatc
      781 ctgcaggtgc tctgtcagga ccacgggctt acgccacaac aggtggtggc gattgcatct
      841 aatcacggcg gacgccctgc cttggtgtcc atcgtagccc aattgtccag gcccgatccc
      901 gcgttggctg cgttaacgaa tgaccatctg gtggcgttgg catgtcttgg tggacgaccc
      961 gcgctcgatg cagtcaaaaa gggtctgcct catgctcccg cattgatcaa aagaaccaac
     1021 cggcggattc ccgagagaac ttcccatcga gtcgcggcta gccccaagaa gaagagaaag
     1081 gtggaggcca gcggttccgg acagtggatg gacgcgaaat cacttacggc atggtcgaga
     1141 acactggtta cgttcaagga cgtgtttgtg gactttacac gtgaggagtg gaaattgctg
     1201 gatactgcgc aacaaattgt gtatcgaaat gtcatgcttg agaattacaa gaacctcgtc
     1261 agtctcggat accagttgac gaaaccggat gtgatcctta ggctcgaaaa gggggaagaa
     1321 ccttggctgg tataaagctg agctcgctga tcagcctcga ctgtgccttc tagttgccag
     1381 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1441 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1501 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     1561 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaacctgcag cccaagcttg
     1621 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1681 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1741 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     1801 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     1861 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     1921 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     1981 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     2041 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2101 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     2161 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2221 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2281 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2341 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2401 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2461 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2521 cttaccgcat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2581 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2641 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2701 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2761 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2821 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2881 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2941 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3001 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3061 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3121 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3181 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3241 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3301 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3361 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3421 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3481 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3541 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3601 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3661 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3721 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3781 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3841 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3901 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3961 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatgaccg
     4021 ctttggaaaa acaaagactg tatttcctgg aaattaatgt ttattcaata aactgtgtat
     4081 tcagctatat tcacatagtg gtgaggctga aatgaggcgg gaagaggcgg ttggggctta
     4141 attatatcaa tttgggtggc cccacagcgc ctccaaggcg ccagtcctgt tttgacaagt
     4201 tgcctctgga agcctctctt ctttttctcc agagtaagcg gaggccaggg gcccccggcc
     4261 tctgcttaat actaaaaaaa acagctgttg tcatagtaat gattgggtgg aaacattcca
     4321 ggcctgggtg gagaggcttt ttgcttcctc ttgcaaaacc acactgacat tccaggcctg
     4381 ggtggagagg ctttttgctt cctcttgcaa aaccacactg ccctctggag ggcagttgcc
     4441 tagcaactaa ttaaaagagg atgtcgcacg gccagctgcg gtcagttagt cacttcctgc
     4501 ttaactgact tgacattttc tattttaaga gtcgggagga aaattactgt gttggaggcc
     4561 ctccgccatc ttctgaagct gaatcgaatt aacttgttta ttgcagctta taatggttac
     4621 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4681 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgata tcatcgtc
//
