LOCUS       sequence-204663-        7760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA3 - pCMV - PhyB NT - GBD, expressing Phytochrome B (PhyB) with 
            Gal Binding Domain (GBD), under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7760)
  AUTHORS   Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Hu VJ, Chao SS, 
            Hsu A, Pham V, Naghavian L, Dozier LE, Patrick GN, Coleman TP
  TITLE     Biosynthesis of Orthogonal Molecules Using Ferredoxin and 
            Ferredoxin-NADP(+) Reductase Systems Enables Genetically Encoded 
            PhyB Optogenetics.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi: 
            10.1021/acssynbio.7b00413. Epub 2018 Jan 24.
  PUBMED    29301067
REFERENCE   2  (bases 1 to 7760)
  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"; doi: 
            "10.1021/acssynbio.7b00413"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "706-717"
COMMENT     Sequence Label: pPKm-105
FEATURES             Location/Qualifiers
     source          1..7760
                     /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             2779..2799
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2809..3249
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             3265..3291
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3313..3331)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3314..3331)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3351..3368)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3357..3581
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3627..4055
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3714..3733)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3924..3945
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4064..4084)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4069..4398
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4249..4384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4311..4330
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4465..5259
                     /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(4519..4538)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5129..5148
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5433..5554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5470..5489)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5524..5543
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5603..5619)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5603..5619)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5616..5638)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5627..5643
                     /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(5651..5657,5658..5675,5676..5681))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5696..5717
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5834..5851)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6005..6593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6085..6104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6764..7555,7556..7624))
                     /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     7387..7406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7625..7729)
                     /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 gcttggtacc
      901 aacatggttt ccggagtcgg gggtagtggc ggtggccgtg gcggtggccg tggcggagaa
      961 gaagaaccgt cgtcaagtca cactcctaat aaccgaagag gaggagaaca agctcaatcg
     1021 tcgggaacga aatctctcag accaagaagc aacactgaat caatgagcaa agcaattcaa
     1081 cagtacaccg tcgacgcaag actccacgcc gttttcgaac aatccggcga atcagggaaa
     1141 tcattcgact actcacaatc actcaaaacg acgacgtacg gttcctctgt acctgagcaa
     1201 cagatcacag cttatctctc tcgaatccag cgaggtggtt acattcagcc tttcggatgt
     1261 atgatcgccg tcgatgaatc cagtttccgg atcatcggtt acagtgaaaa cgccagagaa
     1321 atgttaggga ttatgcctca atctgttcct actcttgaga aacctgagat tctagctatg
     1381 ggaactgatg tgagatcttt gttcacttct tcgagctcga ttctactcga gcgtgctttc
     1441 gttgctcgag agattacctt gttaaatccg gtttggatcc attccaagaa tactggtaaa
     1501 ccgttttacg ccattcttca taggattgat gttggtgttg ttattgattt agagccagct
     1561 agaactgaag atcctgcgct ttctattgct ggtgctgttc aatcgcagaa actcgcggtt
     1621 cgtgcgattt ctcagttaca ggctcttcct ggtggagata ttaagctttt gtgtgacact
     1681 gtcgtggaaa gtgtgaggga cttgactggt tatgatcgtg ttatggttta taagtttcat
     1741 gaagatgagc atggagaagt tgtagctgag agtaaacgag acgatttaga gccttatatt
     1801 ggactgcatt atcctgctac tgatattcct caagcgtcaa ggttcttgtt taagcagaac
     1861 cgtgtccgaa tgatagtaga ttgcaatgcc acacctgttc ttgtggtcca ggacgatagg
     1921 ctaactcagt ctatgtgctt ggttggttct actcttaggg ctcctcatgg ttgtcactct
     1981 cagtatatgg ctaacatggg atctattgcg tctttagcaa tggcggttat aatcaatgga
     2041 aatgaagatg atgggagcaa tgtagctagt ggaagaagct cgatgaggct ttggggtttg
     2101 gttgtttgcc atcacacttc ttctcgctgc ataccgtttc cgctaaggta tgcttgtgag
     2161 tttttgatgc aggctttcgg tttacagtta aacatggaat tgcagttagc tttgcaaatg
     2221 tcagagaaac gcgttttgag aacgcagaca ctgttatgtg atatgcttct gcgtgactcg
     2281 cctgctggaa ttgttacaca gagtcccagt atcatggact tagtgaaatg tgacggtgca
     2341 gcatttcttt accacgggaa gtattacccg ttgggtgttg ctcctagtga agttcagata
     2401 aaagatgttg tggagtggtt gcttgcgaat catgcggatt caaccggatt aagcactgat
     2461 agtttaggcg atgcggggta tcccggtgca gctgcgttag gggatgctgt gtgcggtatg
     2521 gcagttgcat atatcacaaa aagagacttt cttttttggt ttcgatctca cactgcgaaa
     2581 gaaatcaaat ggggaggcgc taagcatcat ccggaggata aagatgatgg gcaacgaatg
     2641 catcctcgtt cgtcctttca ggcttttctt gaagttgtta agagccggag tcagccatgg
     2701 gaaactgcgg aaatggatgc gattcactcg ctccagctta ttctgagaga ctcttttaaa
     2761 gaatctttgc ggccgcaacc taagaaaaag agaaaggtgg gcggaggcat gaagctactg
     2821 tcttctatcg aacaagcatg cgatatttgc cgacttaaaa agctcaagtg ctccaaagaa
     2881 aaaccgaagt gcgccaagtg tctgaagaac aactgggagt gtcgctactc tcccaaaacc
     2941 aaaaggtctc cgctgactag ggcacatctg acagaagtgg aatcaaggct agaaagactg
     3001 gaacagctat ttctactgat ttttcctcga gaagaccttg acatgatttt gaaaatggat
     3061 tctttacagg atataaaagc attgttaaca ggattatttg tacaagataa tgtgaataaa
     3121 gatgccgtca cagatagatt ggcttcagtg gagactgata tgcctctaac attgagacag
     3181 catagaataa gtgcgacatc atcatcggaa gagagtagta acaaaggtca aagacagttg
     3241 actgtatcgc cggaattcat ggcttaccca tacgatgttc cagattacgc ttgataatct
     3301 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     3361 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3421 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3481 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3541 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3601 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3661 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3721 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3781 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3841 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3901 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3961 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4021 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4081 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4141 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4201 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4261 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4321 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4381 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4441 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4501 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4561 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4621 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4681 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4741 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4801 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4861 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4921 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4981 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5041 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5101 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5161 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5221 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5281 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5341 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5401 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5461 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5521 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5581 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5641 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5701 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5761 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5821 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5881 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5941 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6001 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6061 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6121 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6181 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6241 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6301 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6361 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6421 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6481 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6541 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6601 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6661 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6721 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6781 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6841 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6901 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6961 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7021 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7081 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7141 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7201 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7261 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7321 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7381 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7441 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7501 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7561 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7621 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7681 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7741 gaaaagtgcc acctgacgtc
//
