LOCUS       sequence-204641-        7172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA3 - pCMV - MTAD - PIF3, expresses Phytochrome interacting 
            factor 3 (PIF3) and minimal transactivation domain (MTAD), under CMV
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7172)
  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 7172)
  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-112
FEATURES             Location/Qualifiers
     source          1..7172
                     /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             1093..1104
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(2714..2732)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2715..2732)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2752..2769)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2758..2982
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3028..3456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3115..3134)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3325..3346
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3465..3485)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3470..3799
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3650..3785
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3712..3731
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3866..4660
                     /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(3920..3939)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4530..4549
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4834..4955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4871..4890)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4925..4944
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5004..5020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5004..5020)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5017..5039)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5028..5044
                     /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(5052..5058,5059..5076,5077..5082))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5097..5118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5235..5252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5417..6005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5497..5516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6176..6967,6968..7036))
                     /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     6799..6818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7037..7141)
                     /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 gagctcggat ccatgggatc tcctaggttt gatactgacc tgttcgatac cgacttgttt
      961 gatactgacc tttttgatac tgacctgttc gataccgact tgtttgatac tgaccttttt
     1021 gatactgacc tgttcgatac cgacttgttt gatactgacc tttttgatac tgacctgttc
     1081 gataccgact tgatcgaggg aaggggtggc gaattcatgc ctctgtttga gcttttcagg
     1141 ctcaccaaag ctaagcttga atctgctcaa gacaggaacc cttctccacc tgtagatgaa
     1201 gttgtggagc tggtgtggga aaatggtcag atatcaactc aaagtcagtc aagtagatcg
     1261 aggaacattc ctccaccaca agcaaactct tctagagcta gagagattgg aaatggctca
     1321 aagacgacta tggtggacga gatccctatg tcagtgccat cactaatgac gggtttgagt
     1381 caagacgatg actttgttcc atggttgaat catcatccct cccttgatgg atattgctct
     1441 gatttcttgc gtgatgtgtc gtctcctgtt actgtcaacg agcaagagag tgatatggcg
     1501 gtaaaccaaa ctgctttccc gttgtttcag agaagaaagg atggcaatga atcagctcct
     1561 gctgcttctt cgtcgcagta taacggtttc caatcgcatt ctctgtatgg aagtgataga
     1621 gctagagatc ttcctagcca acaaaccaat ccggatcggt ttactcagac gcaggaacca
     1681 ctaattacta gtaacaagcc tagtttggtc aacttttcac atttcttacg ccctgcaact
     1741 tttgcgaaga ctactaataa taaccttcat gacactaaag aaaagagtcc tcaaagcccg
     1801 ccaaatgtgt ttcagaccag agttcttgga gctaaagact ctgaagataa ggttcttaac
     1861 gagtctgttg cttctgctac gcctaaagat aaccaaaagg cttgcctaat atcagaggac
     1921 tcatgtagaa aagaccaaga gagtgaaaaa gcagttgtat gttcttctgt tggctcgggt
     1981 aatagtctcg atggcccatc cgaaagtcct tcactttctt taaagagaaa gcattcgaat
     2041 attcaagaca ttgactgtca tagtgaagat gtggaagaag aatcaggaga tggaagaaag
     2101 gaagcaggtc catctcgaac gggtttgggt tcaaagagaa gccgctctgc agaagtgcat
     2161 aatctgtctg aaaggagacg gcgtgatagg atcaacgaga agatgcgtgc cctgcaagaa
     2221 ctcattccaa actgtaacaa ggtggacaaa gcttcgatgc tagatgaagc catcgagtat
     2281 ctcaagtcac tccaacttca agtgcagatc atgtcaatgg cgtctggtta ctatctgcca
     2341 ccggcggtta tgttcccacc gggtatgggg cattacccgg cagcagctgc tgcaatggca
     2401 atgggtatgg gaatgcctta tgcaatgggc ttgcctgatt tgagccgtgg tggttcatcg
     2461 gttaaccacg gaccacagtt ccaagtctcg gggatgcaac aacaaccagt ggcgatgggt
     2521 attccacgtg tctctggtgg tggtatcttt gccggttctt cgacgattgg caatggctcg
     2581 actagagatt tatctggttc taaagatcaa acaacgacga ataacaacag taacttgaaa
     2641 ccaataaaga gaaaacaggg gtcttctgat cagttttgtg gatcgtcgta agcatgcatc
     2701 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2761 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2821 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2881 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2941 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3001 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3061 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3121 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3181 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3241 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3301 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3361 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3421 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3481 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3541 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3601 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3661 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3721 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3781 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3841 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3901 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3961 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4021 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4081 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4141 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4201 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4261 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4321 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4381 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4441 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4501 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4561 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4621 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4681 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4741 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4801 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4861 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4921 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4981 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5041 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5101 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5161 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5221 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5281 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5341 aagaacatgt gagcaaaagg ccagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5401 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5461 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5521 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5581 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5641 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5701 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5761 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5821 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5881 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5941 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6001 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6061 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6121 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6181 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6241 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6301 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6361 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6421 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6481 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6541 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6601 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6661 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6721 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6781 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6841 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6901 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6961 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7021 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7081 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7141 gcacatttcc ccgaaaagtg ccacctgacg tc
//
