LOCUS       Exported                6300 bp ds-DNA     circular SYN 13-MAY-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 6300)
  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 6300)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6300
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..658
                     /label=chicken beta-actin promoter
     intron          659..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1731..1740
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1740..1805
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1812..1832
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2615..2635
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2690..2884
                     /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(2907..2924)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2913..3137
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3184..3200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3184..3200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3197..3219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3208..3224
                     /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(3232..3262)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3277..3298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3357..3552
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3403..3538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3465..3484
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3558..3692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3760..3777)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3931..4519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4011..4030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4690..5550)
                     /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     5313..5332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6153..6287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg
      721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct
      841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc
     1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1381 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1441 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     1501 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttgt tgtgctgtct catcattttg gcaaagaatt cctcgaggcc gccaccatgg
     1741 actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg
     1801 acaagatggc ccccaagaag aagaggaagg tgggcattca ccgcggggta cctatggtgg
     1861 acttgaggac actcggttat tcgcaacagc aacaggagaa aatcaagcct aaggtcagga
     1921 gcaccgtcgc gcaacaccac gaggcgcttg tggggcatgg cttcactcat gcgcatattg
     1981 tcgcgctttc acagcaccct gcggcgcttg ggacggtggc tgtcaaatac caagatatga
     2041 ttgcggccct gcccgaagcc acgcacgagg caattgtagg ggtcggtaaa cagtggtcgg
     2101 gagcgcgagc acttgaggcg ctgctgactg tggcgggtga gcttaggggg cctccgctcc
     2161 agctcgacac cgggcagctg ctgaagatcg cgaagagagg gggagtaaca gcggtagagg
     2221 cagtgcacgc ctggcgcaat gcgctcaccg gggccccctt gaacctgcag gatacgttta
     2281 tcagtgataa agtgtcaagc atgacaaagt tgcagccgaa tacagtggta tcctgcaggt
     2341 gctctgtcag gaccacgggc ttacgccaca acaggtggtg gcgattgcat ctaatcacgg
     2401 cggacgccct gccttggtgt ccatcgtagc ccaattgtcc aggcccgatc ccgcgttggc
     2461 tgcgttaacg aatgaccatc tggtggcgtt ggcatgtctt ggtggacgac ccgcgctcga
     2521 tgcagtcaaa aagggtctgc ctcatgctcc cgcattgatc aaaagaacca accggcggat
     2581 tcccgagaga acttcccatc gagtcgcggc tagccccaag aagaagagaa aggtggaggc
     2641 cagcggttcc ggacagtgga tggacgcgaa atcacttacg gcatggtcga gaacactggt
     2701 tacgttcaag gacgtgtttg tggactttac acgtgaggag tggaaattgc tggatactgc
     2761 gcaacaaatt gtgtatcgaa atgtcatgct tgagaattac aagaacctcg tcagtctcgg
     2821 ataccagttg acgaaaccgg atgtgatcct taggctcgaa aagggggaag aaccttggct
     2881 ggtataaagc tgagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2941 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3001 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3061 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3121 tgcggtgggc tctatggctt ctgaggcgga aagaacctgc agcccaagct tggcgtaatc
     3181 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3241 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3301 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
     3361 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3421 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3481 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3541 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3601 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3661 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
     3721 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3781 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3841 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3901 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3961 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4021 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgc
     4081 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     4141 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4201 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4261 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4321 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4381 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4441 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4501 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4561 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4621 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4681 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4741 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4801 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4861 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4921 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4981 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5041 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5101 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5161 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5221 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5281 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5341 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5401 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5461 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5521 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatgac cgctttggaa
     5581 aaacaaagac tgtatttcct ggaaattaat gtttattcaa taaactgtgt attcagctat
     5641 atcacatagt ggtgaggctg aaatgaggcg ggaagaggcg gttggggctt aattatatca
     5701 atttgggtgg ccccacagcg cctccaaggc gccagtcctg ttttgacaag ttgcctctgg
     5761 aagcctctac aatgcctctc ttctttttct ccagagtaag cggaggccag gggcccccgg
     5821 cctctgctta atactaaaaa aaacagctgt tgtcatagta atgattgggt ggaaacattc
     5881 caggcctggg tggagaggct ttttgcttcc tcttgcaaaa ccacactgac attccaggcc
     5941 tgggtggaga ggctttttgc ttcctcttgc aaaaccacac tgccctctgg agggcagttg
     6001 cctagcaact aactaaaaga ggatgtcgca cggccagctg cggtcagtta gtcacttcct
     6061 gcttaactga cttgacattt tctattttaa gagtcgggag gaaaattact gtgttggagg
     6121 ccctccgcca tcttctgaag ctgaatcgaa ttaacttgtt tattgcagct tataatggtt
     6181 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6241 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga tatcatcgtc
//
