LOCUS       sequence-204672-        6647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pcDNA3 - CMVmin  - 5X UAS - pFR - CMVmin  Luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6647)
  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 6647)
  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-202
FEATURES             Location/Qualifiers
     source          1..6647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     621..640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(859..963)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1038..1057)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    1235..1329
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     primer_bind     1373..1397
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1501..1510
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1507..3159
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTVLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1573..1591)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(3194..3212)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3195..3212)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3232..3249)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3238..3462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3508..3936
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3595..3614)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3805..3826
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3945..3965)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3950..4279
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4130..4265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4192..4211
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4346..5140
                     /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(4400..4419)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5010..5029
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5314..5435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5351..5370)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5405..5424
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5484..5500)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5484..5500)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5497..5519)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5508..5524
                     /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(5532..5538,5539..5556,5557..5562))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5577..5598
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5715..5732)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5886..6474)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5966..5985)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6645..6647,1..789,790..858))
                     /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 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
       61 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      121 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      181 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      241 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      301 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      361 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      421 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      481 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      541 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      601 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      661 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      721 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      781 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      841 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      901 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      961 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     1021 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     1081 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     1141 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     1201 cgcgaaagct tgcatgcgag tactgtcctc cgagcggagt actgtcctcc gagcggagta
     1261 ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag cggagtactg
     1321 tcctccgagc ggagactcta gagggtatat aatggatccc ctatataagc agagctcgtt
     1381 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
     1441 accgggaccg atccagcctg gtaccgagct cgaattccag cttggcattc cggtactgcc
     1501 gccaccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta tcctctagag
     1561 gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga
     1621 acaattgctt ttacagatgc acatatcgag gtgaacatca cgtacgcgga atacttcgaa
     1681 atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc
     1741 gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc
     1801 ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg
     1861 aacatttcgc agcctaccgt agtgtttgtt tccaaaaagg ggttgcaaaa aattttgaac
     1921 gtgcaaaaaa aattaccaat aatccagaaa attattatca tggattctaa aacggattac
     1981 cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa
     2041 tacgattttg taccagagtc ctttgatcgt gacaaaacaa ttgcactgat aatgaattcc
     2101 tctggatcta ctgtgttacc taagggtgtg gcccttccgc atagaactgc ctgcgtcaga
     2161 ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta
     2221 agtgttgttc cattccatca cggttttgga atgtttacta cactcggata tttgatatgt
     2281 ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttttacg atcccttcag
     2341 gattacaaaa ttcaaagtgc gttgctagta ccaaccctat tttcattctt cgccaaaagc
     2401 actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg gggcgcacct
     2461 ctttcgaaag aagtcgggga agcggttgca aaacgcttcc atcttccagg gatacgacaa
     2521 ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa
     2581 ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc
     2641 gggaaaacgc tgggcgttaa tcagagaggc gaattatgtg tcagaggacc tatgattatg
     2701 tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta
     2761 cattctggag acatagctta ctgggacgaa gacgaacact tcttcatagt tgaccgcttg
     2821 aagtctttaa ttaaatacaa aggatatcag gtggcccccg ctgaattgga atcgatattg
     2881 ttacaacacc ccaacatctt cgacgcgggc gtggcaggtc ttcccgacga tgacgccggt
     2941 gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc
     3001 gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt
     3061 gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc
     3121 ctcataaagg ccaagaaggg cggaaagtcc aaattgtaag cggccgctcg agcatgcatc
     3181 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     3241 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3301 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3361 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3421 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3481 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3541 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3601 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3661 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3721 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3781 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3841 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3901 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3961 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4081 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4141 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4201 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4261 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4321 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4381 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4441 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4501 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4561 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4621 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4681 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4741 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4801 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4861 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4921 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4981 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5041 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5101 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5161 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5221 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5281 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5341 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5401 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5461 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5521 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5581 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5641 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5701 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5761 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5821 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5881 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5941 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6001 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6061 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6121 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6181 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6241 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6301 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6361 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6421 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6481 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6541 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6601 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagtta
//
