LOCUS       sequence-59235-e        6770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Activation Domain: NF-kB; RVD of C-terminal 0.5 TALE repeat: NI 
            (binds A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6770)
  AUTHORS   Maeder ML, Linder SJ, Reyon D, Angstman JF, Fu Y, Sander JD, Joung 
            JK
  TITLE     Robust, synergistic regulation of human gene expression using TALE 
            activators.
  JOURNAL   Nat Methods. 2013 Feb 10. doi: 10.1038/nmeth.2366.
  PUBMED    23396285
REFERENCE   2  (bases 1 to 6770)
  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: "Nat 
            Methods. 2013 Feb 10. doi: 10.1038/nmeth.2366."
COMMENT     Sequence Label: MLM2581
FEATURES             Location/Qualifiers
     source          1..6770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     331..350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(569..673)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(752..771)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        927..2105
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1158..2096
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2053..2073
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2113..2122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2125..2145
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3397..3768
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     primer_bind     complement(3810..3829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3864..3883
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3920..3940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3925..4254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4105..4240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4264..4863
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(4264..4283)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4760..4780
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4925..4966
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4976..4993
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5016..5033)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5022..5246
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5317..5333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5317..5333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5330..5352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5341..5357
                     /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(5365..5371,5372..5389,5390..5395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5410..5431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5548..5565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5719..6307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5799..5818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6478..6770,1..499,500..568))
                     /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"
ORIGIN
        1 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
       61 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
      121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
      181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
      241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
      301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
      361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
      421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
      481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
      541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
      601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
      661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
      721 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
      781 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      841 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      901 gttttgcgct gcttcgctag tcgtgaggct ccggtgcccg tcagtgggca gagcgcacat
      961 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     1021 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     1081 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     1141 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     1201 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     1261 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     1321 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     1381 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     1441 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     1501 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     1561 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     1621 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     1681 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     1741 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     1801 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1861 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1921 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1981 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     2041 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     2101 cgtgaagcgg ccgccaccat ggcccccaag aagaagagga aggtgggcat tcaccgcggg
     2161 gtacctatgg tggacttgag gacactcggt tattcgcaac agcaacagga gaaaatcaag
     2221 cctaaggtca ggagcaccgt cgcgcaacac cacgaggcgc ttgtggggca tggcttcact
     2281 catgcgcata ttgtcgcgct ttcacagcac cctgcggcgc ttgggacggt ggctgtcaaa
     2341 taccaagata tgattgcggc cctgcccgaa gccacgcacg aggcaattgt aggggtcggt
     2401 aaacagtggt cgggagcgcg agcacttgag gcgctgctga ctgtggcggg tgagcttagg
     2461 gggcctccgc tccagctcga caccgggcag ctgctgaaga tcgcgaagag agggggagta
     2521 acagcggtag aggcagtgca cgcctggcgc aatgcgctca ccggggcccc cttgaacaga
     2581 gacgatctag agctagacgt ctcgctgaca cccgaacagg tggtcgccat tgcttctaac
     2641 atcggaggac ggccagcctt ggagagcatt gtggcccagc tgtccaggcc ggaccctgcc
     2701 ctggccgcgt taaccaacga ccacctcgtc gccttggcct gcctcggcgg acgtcctgcg
     2761 ctggatgcag tgaaaaaggg attgccgcac gcgccggcct tgatcaaaag aaccaatcgc
     2821 cgtattcccg aacgcacatc ccatcgcgtt gccgaccacg cgcaagtggt tcgcgtgctg
     2881 ggttttttcc agtgccactc ccacccagcg caagcatttg atgacgccat gacgcagttc
     2941 gggatgagcg gaggaggatc ccccatggag ttccagtacc tgccagatac agacgatcgt
     3001 caccggattg aggagaaacg taaaaggaca tatgagacct tcaagagcat catgaagaag
     3061 agtcctttca gcggacccac cgacccccgg cctccacctc gacgcattgc tgtgccttcc
     3121 cgcagctcag cttctgtccc caagccagca ccccagccct atccctttac gtcatccctg
     3181 agcaccatca actatgatga gtttcccacc atggtgtttc cttctgggca gatcagccag
     3241 gcctcggcct tggccccggc ccctccccaa gtcctgcccc aggctccagc ccctgcccct
     3301 gctccagcca tggtatcagc tctggcccag gccccagccc ctgtcccagt cctagcccca
     3361 ggccctcctc aggctgtggc cccacctgcc cccaagccca cccaggctgg ggaaggaacg
     3421 ctgtcagagg ccctgctgca gctgcagttt gatgatgaag acctgggggc cttgcttggc
     3481 aacagcacag acccagctgt gttcacagac ctggcatccg tcgataactc cgagtttcag
     3541 cagctgctga accagggcat acctgtggcc ccccacacaa ctgagcccat gctgatggag
     3601 taccctgagg ctataactcg cctagtgaca ggggcccaga ggccccccga cccagctcct
     3661 gctccactgg gggccccggg gctccccaat ggcctccttt caggagatga agacttctcc
     3721 tccattgcgg acatggactt ctcagccctg ctgagtcaga tcagctctta attaattgtt
     3781 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3841 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatgaatt
     3901 ccgattaagt cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4141 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcttct
     4201 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     4261 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     4321 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     4381 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     4441 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     4501 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     4561 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     4621 gagcccgcgt ggttcctggc caccgtcggc gtgtcgcccg accaccaggg caagggtctg
     4681 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     4741 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     4801 gccgacgtcg agtgcccgaa ggaccgcgcg acctggtgca tgacccgcaa gcccggtgcc
     4861 tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg
     4921 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     4981 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     5041 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5101 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     5161 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     5221 tgctggggat gcggtgggct ccatggcttc tgaggcggaa agaaccagct ggggctcgat
     5281 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5341 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     5401 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5461 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5521 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5581 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5641 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5881 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     5941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6121 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
//
