LOCUS       sequence-204685-        7476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA3 - CMVmin - PhyB - MTAD, expresses PhyB and MTAD, under CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7476)
  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 7476)
  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-195
FEATURES             Location/Qualifiers
     source          1..7476
                     /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             2950..2961
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2981..3001
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(3029..3047)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3030..3047)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3067..3084)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3073..3297
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3343..3771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3430..3449)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3640..3661
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3780..3800)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3785..4114
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3965..4100
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4027..4046
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4181..4975
                     /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(4235..4254)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4845..4864
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5149..5270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5186..5205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5240..5259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5319..5335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5319..5335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5332..5354)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5343..5359
                     /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(5367..5373,5374..5391,5392..5397))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5412..5433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5550..5567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5721..6309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5801..5820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6480..7271,7272..7340))
                     /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     7103..7122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7341..7445)
                     /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 atggtttccg gagtcggggg tagtggcggt ggccgtggcg gtggccgtgg cggagaagaa
      961 gaaccgtcgt caagtcacac tcctaataac cgaagaggag gagaacaagc tcaatcgtcg
     1021 ggaacgaaat ctctcagacc aagaagcaac actgaatcaa tgagcaaagc aattcaacag
     1081 tacaccgtcg acgcaagact ccacgccgtt ttcgaacaat ccggcgaatc agggaaatca
     1141 ttcgactact cacaatcact caaaacgacg acgtacggtt cctctgtacc tgagcaacag
     1201 atcacagctt atctctctcg aatccagcga ggtggttaca ttcagccttt cggatgtatg
     1261 atcgccgtcg atgaatccag tttccggatc atcggttaca gtgaaaacgc cagagaaatg
     1321 ttagggatta tgcctcaatc tgttcctact cttgagaaac ctgagattct agctatggga
     1381 actgatgtga gatctttgtt cacttcttcg agctcgattc tactcgagcg tgctttcgtt
     1441 gctcgagaga ttaccttgtt aaatccggtt tggatacatt ccaagaatac tggtaaaccg
     1501 ttttacgcca ttcttcatag gattgatgtt ggtgttgtta ttgatttaga gccagctaga
     1561 actgaagatc ctgcgctttc tattgctggt gctgttcaat cgcagaaact cgcggttcgt
     1621 gcgatttctc agttacaggc tcttcctggt ggagatatta agcttttgtg tgacactgtc
     1681 gtggaaagtg tgagggactt gactggttat gatcgtgtta tggtttataa gtttcatgaa
     1741 gatgagcatg gagaagttgt agctgagagt aaacgagacg atttagagcc ttatattgga
     1801 ctgcattatc ctgctactga tattcctcaa gcgtcaaggt tcttgtttaa gcagaaccgt
     1861 gtccgaatga tagtagattg caatgccaca cctgttcttg tggtccagga cgataggcta
     1921 actcagtcta tgtgcttggt tggttctact cttagggctc ctcatggttg tcactctcag
     1981 tatatggcta acatgggatc tattgcgtct ttagcaatgg cggttataat caatggaaat
     2041 gaagatgatg ggagcaatgt agctagtgga agaagctcga tgaggctttg gggtttggtt
     2101 gtttgccatc acacttcttc tcgctgcata ccgtttccgc taaggtatgc ttgtgagttt
     2161 ttgatgcagg ctttcggttt acagttaaac atggaattgc agttagcttt gcaaatgtca
     2221 gagaaacgcg ttttgagaac gcagacactg ttatgtgata tgcttctgcg tgactcgcct
     2281 gctggaattg ttacacagag tcccagtatc atggacttag tgaaatgtga cggtgcagca
     2341 tttctttacc acgggaagta ttacccgttg ggtgttgctc ctagtgaagt tcagataaaa
     2401 gatgttgtgg agtggttgct tgcgaatcat gcggattcaa ccggattaag cactgatagt
     2461 ttaggcgatg cggggtatcc cggtgcagct gcgttagggg atgctgtgtg cggtatggca
     2521 gttgcatata tcacaaaaag agactttctt ttttggtttc gatctcacac tgcgaaagaa
     2581 atcaaatggg gaggcgctaa gcatcatccg gaggataaag atgatgggca acgaatgcat
     2641 cctcgttcgt cctttcaggc ttttcttgaa gttgttaaga gccggagtca gccatgggaa
     2701 actgcggaaa tggatgcgat tcactcgctc cagcttattc tgagagactc ttttaaagaa
     2761 tctggatcca tgggatctcc taggtttgat actgacctgt tcgataccga cttgtttgat
     2821 actgaccttt ttgatactga cctgttcgat accgacttgt ttgatactga cctttttgat
     2881 actgacctgt tcgataccga cttgtttgat actgaccttt ttgatactga cctgttcgat
     2941 accgacttga tcgagggaag gggtggcgaa ttaattcatg cctaagaaaa agagaaaggt
     3001 gggcggaggc gctagtagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     3061 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3121 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3181 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3241 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3301 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3361 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3421 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3481 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3541 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3601 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3661 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3721 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3781 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3841 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3901 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3961 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4021 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4081 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4141 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4201 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4261 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4321 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4381 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4441 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4501 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4561 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4621 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4681 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4741 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4801 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4861 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4921 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4981 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5041 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5101 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5161 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5221 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5281 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5341 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5401 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5461 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5521 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5581 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5641 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5701 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5761 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5821 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5881 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5941 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6061 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6121 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6181 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6241 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6301 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6361 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6421 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6481 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6541 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6601 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6661 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6721 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6781 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6841 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6901 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6961 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7021 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7081 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7141 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7201 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7261 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7321 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7381 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7441 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
