LOCUS       sequence-310671-        6819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LDB-14 fused with a C-terminal p65 transcriptional 
            activation domain in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6819)
  AUTHORS   Huang Z, Li Z, Zhang X, Kang S, Dong R, Sun L, Fu X, Vaisar D, 
            Watanabe K, Gu L
  TITLE     Creating Red Light-Switchable Protein Dimerization Systems as 
            Genetically Encoded Actuators with High Specificity.
  JOURNAL   ACS Synth Biol. 2020 Nov 12. doi: 10.1021/acssynbio.0c00397.
  PUBMED    33179507
REFERENCE   2  (bases 1 to 6819)
  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: "ACS Synth 
            Biol. 2020 Nov 12. doi: 10.1021/acssynbio.0c00397."
COMMENT     Sequence Label: LDB-14-p65
FEATURES             Location/Qualifiers
     source          1..6819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             910..930
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             946..966
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1876..2247
                     /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"
     CDS             2317..2358
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2368..2385
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2408..2425)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2414..2638
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2684..3112
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2771..2790)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2981..3002
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3121..3141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3126..3455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3306..3441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3368..3387
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3522..4316
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3576..3595)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4186..4205
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4492..4613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4529..4548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4583..4602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4662..4678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4662..4678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4675..4697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4686..4702
                     /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(4710..4716,4717..4734,4735..4740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4755..4776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4893..4910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5064..5652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5144..5163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5823..6614,6615..6683))
                     /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     6446..6465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6684..6788)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagctatgc ccaagaagaa gcgcaaggtg gaagctagcg cttccccgaa gaaaaagcgg
      961 aaagtcgagg cctccgcatc tccaaaaaaa aaaagcaagg ttgaagcatc tggatccgga
     1021 ggcggttccg gtggcggttc tgaagttcag ctgcaggcaa gcggtggtgg ttttgttcag
     1081 cctggtggta gcctgcgtct gagctgtgca gccagcggta ccacctctcg ttgggaatct
     1141 atgggctggt ttcgccaggc accgggtaaa gaacgtgaat ttgttagcgc aatcagctgg
     1201 cagaataatt ctgttccata ttatgccgat agcgtgaaag gtcgctttac cattagccgt
     1261 gataatagca aaaataccgt ttacctgcag atgaatagtc tgcgtgcaga agataccgca
     1321 acctattatt gtgcagcaca gcataacttt ctgggtcatc gttattgggg tcagggcacc
     1381 caggttaccg ttagcagcgg taccggagga agtggcagct ctggcggcag tggagggtct
     1441 ggtggcagcg gacagtacct gccagataca gacgatcgtc accggattga ggagaaacgt
     1501 aaaaggacat atgagacctt caagagcatc atgaagaaga gtcctttcag cggacccacc
     1561 gacccccggc ctccacctcg acgcattgct gtgccttccc gcagctcagc ttctgtcccc
     1621 aagccagcac cccagcccta tccctttacg tcatccctga gcaccatcaa ctatgatgag
     1681 tttcccacca tggtgtttcc ttctgggcag atcagccagg cctcggcctt ggccccggcc
     1741 cctccccaag tcctgcccca ggctccagcc cctgcccctg ctccagccat ggtatcagct
     1801 ctggcccagg ccccagcccc tgtcccagtc ctagccccag gccctcctca ggctgtggcc
     1861 ccacctgccc ccaagcccac ccaggctggg gaaggaacgc tgtcagaggc cctgctgcag
     1921 ctgcagtttg atgatgaaga cctgggggcc ttgcttggca acagcacaga cccagctgtg
     1981 ttcacagacc tggcatccgt cgacaactcc gagtttcagc agctgctgaa ccagggcata
     2041 cctgtggccc cccacacaac tgagcccatg ctgatggagt accctgaggc tataactcgc
     2101 ctagtgacag gggcccagag gccccccgac ccagctcctg ctccactggg ggccccgggg
     2161 ctccccaatg gcctcctttc aggagatgaa gacttctcct ccattgcgga catggacttc
     2221 tcagccctgc tgagtcagat cagctccgga ggaagtggca gctctggcgg cagtggagaa
     2281 ttcagtagct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
     2341 ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc
     2401 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2461 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2521 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2581 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2641 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2881 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2941 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3001 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3061 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3121 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3181 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3301 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3361 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3421 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3481 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3541 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3601 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3661 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3721 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3781 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3841 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3901 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3961 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4021 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4081 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4141 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4201 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4261 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4321 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4381 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4441 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4501 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4561 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4621 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4681 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4741 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4801 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4861 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4921 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4981 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5041 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5101 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5161 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5221 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5281 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5341 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5401 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5461 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5521 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5581 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5701 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5761 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5821 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5881 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5941 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6001 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6061 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6121 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6181 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6241 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6301 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6361 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6421 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6481 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6541 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6601 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6661 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6721 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6781 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
