LOCUS       Exported                7341 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Destination vector for TALE-VP160 construction.
ACCESSION   .
VERSION     .
KEYWORDS    pTALE-STARVP160-T
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7341)
  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 7341)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7341
                     /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             2678..3061
                     /codon_start=1
                     /product="10 tandem repeats of the minimal activation 
                     domain of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP160
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGS
                     DALDDFDLDMLGSDALDDFDLDML"
     CDS             3083..3139
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3140..3850
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3285..3303)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3553..3572
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3948..3965)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3954..4178
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4225..4241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4225..4241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4238..4260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4249..4265
                     /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(4273..4303)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4318..4339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4398..4593
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4444..4579
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4506..4525
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4599..4733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4801..4818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4972..5560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5052..5071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5731..6591)
                     /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     6354..6373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    7194..7328
                     /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 ctaatggagg
     2401 cggacgccct gccttggagt 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 gacggcgggc tcgcgccgac gcgctggacg atttcgatct
     2701 cgacatgctg ggttctgatg ccctcgatga ctttgacctg gatatgttgg gaagcgacgc
     2761 attggatgac tttgatctgg acatgctcgg ctccgatgct ctggacgatt tcgatctcga
     2821 tatgttaggg tcagacgcac tggatgattt cgaccttgat atgttgggaa gcgatgccct
     2881 tgatgatttc gacctggaca tgctcggcag cgacgccctg gacgatttcg atctggacat
     2941 gctggggtcc gatgccttgg atgattttga cttggatatg ctggggagtg atgccctgga
     3001 cgactttgac ctggacatgc tgggctccga tgcgctcgat gacttcgatt tggatatgtt
     3061 gattaactct agaggcagtg gagccactaa cttctccctg ttgaaacaag caggggatgt
     3121 cgaagagaat cccgggccaa tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
     3181 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     3241 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     3301 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     3361 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     3421 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     3481 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     3541 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     3601 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     3661 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     3721 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     3781 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     3841 gtacaagtaa cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca
     3901 tgcatcgatg atctagagct ccgtctcaag ctgagctcgc tgatcagcct cgactgtgcc
     3961 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4021 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4081 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4141 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaacctg
     4201 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4261 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4321 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4381 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4441 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4501 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4561 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     4621 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4681 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     4741 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4801 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4861 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4921 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4981 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5041 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5101 tccgaccctg ccgcttaccg catacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5161 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5221 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5281 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5341 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5401 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5461 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5521 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5581 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5641 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5701 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5761 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5821 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5881 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5941 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6001 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6061 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6121 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6181 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6241 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6301 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6361 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6421 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6481 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6541 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6601 ttttcaatga ccgctttgga aaaacaaaga ctgtatttcc tggaaattaa tgtttattca
     6661 ataaactgtg tattcagcta tatcacatag tggtgaggct gaaatgaggc gggaagaggc
     6721 ggttggggct taattatatc aatttgggtg gccccacagc gcctccaagg cgccagtcct
     6781 gttttgacaa gttgcctctg gaagcctcta caatgcctct cttctttttc tccagagtaa
     6841 gcggaggcca ggggcccccg gcctctgctt aatactaaaa aaaacagctg ttgtcatagt
     6901 aatgattggg tggaaacatt ccaggcctgg gtggagaggc tttttgcttc ctcttgcaaa
     6961 accacactga cattccaggc ctgggtggag aggctttttg cttcctcttg caaaaccaca
     7021 ctgccctctg gagggcagtt gcctagcaac taactaaaag aggatgtcgc acggccagct
     7081 gcggtcagtt agtcacttcc tgcttaactg acttgacatt ttctatttta agagtcggga
     7141 ggaaaattac tgtgttggag gccctccgcc atcttctgaa gctgaatcga attaacttgt
     7201 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     7261 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     7321 tctggatctg atatcatcgt c
//
