LOCUS       sequence-267559-        7176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of D. melanogaster (Dm) cytochrome with V5 and 
            His tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7176)
  AUTHORS   Ozturk N, Selby CP, Song SH, Ye R, Tan C, Kao YT, Zhong D, Sancar A
  TITLE     Comparative photochemistry of animal type 1 and type 4 
            cryptochromes.
  JOURNAL   Biochemistry. 2009 Sep 15;48(36):8585-93. doi: 10.1021/bi901043s.
  PUBMED    19663499
REFERENCE   2  (bases 1 to 7176)
  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"; journalName: 
            "Biochemistry"; date: "2009-09-15- 15"; volume: "48"; issue: "36"; 
            pages: "8585-93"
COMMENT     Sequence Label: pcDNA3-dCry-V5His
FEATURES             Location/Qualifiers
     source          1..7176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        111..490
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        491..694
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     645..665
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     739..758
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        739..757
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2482..2523
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2533..2550
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2605..2623)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2606..2623)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2643..2660)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2649..2873
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2919..3347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3006..3025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3216..3237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3356..3376)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3361..3690
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3541..3676
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3603..3622
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3757..4551
                     /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(3811..3830)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4421..4440
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4725..4846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4762..4781)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4816..4835
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4895..4911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4895..4911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4908..4930)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4919..4935
                     /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(4943..4949,4950..4967,4968..4973))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4988..5009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5126..5143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5297..5885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5377..5396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6056..6847,6848..6916))
                     /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     6679..6698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6917..7021)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7096..7115)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
       61 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
      121 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      181 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      241 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      301 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      361 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      421 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      481 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      541 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      601 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      661 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
      721 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc
      781 tcggatccac tagtaacggc cgccagtgtg ctggaattca ccatggccac gcgaggggcg
      841 aatgtgattt ggtttcgcca tggattgcgc ctccatgata atcccgctct attggccgcc
      901 ctcgccgata aggatcaggg tatagcccta attcccgttt tcatattcga tggagagagt
      961 gcaggtacca agaatgtggg ttacaatcgg atgcgtttcc tcctggactc gttgcaggac
     1021 atcgatgatc agctacaggc ggcaactgat ggacgtggac gcctcctggt cttcgagggc
     1081 gaaccggctt atatcttccg ccggctacat gagcaagtgc gtctgcacag gatttgcata
     1141 gagcaggact gcgagccaat ttggaatgag cgcgatgaaa gcatccgttc tctatgtcgg
     1201 gagctgaata tcgactttgt cgagaaggta tcacacacgc tttgggatcc gcaattggtg
     1261 attgagacca atggtggcat tccaccgctg acctaccaaa tgttcctgca cacggtgcaa
     1321 attattgggc ttccaccgcg tcccaccgcc gatgctcgac tagaagacgc cacctttgtc
     1381 gagctggacc ccgagttctg ccgaagtctt aagttgttcg agcagctgcc cacgccggag
     1441 cacttcaatg tgtatggaga caacatgggc ttcctggcca agattaactg gcgcggcgga
     1501 gaaacacagg ccttacttct gttggatgag cgtcttaaag tggagcagca tgcgtttgag
     1561 cgtggatttt atctgcccaa ccaggcactg cccaatatcc acgactcgcc aaaatcgatg
     1621 agcgcccatc tgcgctttgg ttgcctttcg gtacgtcgct tctactggag cgtccacgat
     1681 ctcttcaaga atgtccagtt gcgcgcctgt gtgcggggcg ttcagatgac tggcggcgcg
     1741 cacatcacgg gacagttgat ctggcgagag tacttctaca ccatgtcggt gaacaatcca
     1801 aactacgatc gcatggaggg caatgacatc tgcctgagca tcccgtgggc taagccgaac
     1861 gaaaatctcc tgcagagctg gcgtttaggc caaacgggat tcccgctcat cgacggcgcc
     1921 atgcgacaac tcctggccga gggatggctc caccatacgc tgcgcaacac cgtggccacc
     1981 tttctcacgc gcggcggttt gtggcagagc tgggagcatg gactgcagca ctttctgaag
     2041 tatctgctgg atgcggattg gtcggtctgc gctggcaact ggatgtgggt atccagctcg
     2101 gcgtttgaaa ggctgctgga ctcctccctg gtcacctgcc cggtggcatt ggccaagcga
     2161 cttgatccgg atggcaccta catcaagcag tacgtcccgg agttgatgaa tgtgcccaag
     2221 gaatttgttc acgagccctg gcgaatgtct gccgagcagc aggagcagta cgagtgcctg
     2281 atcggagtcc attatccgga gcggatcatt gatttgtcca tggccgtaaa gcgaaacatg
     2341 ctggccatga agtctctccg aaattcgctg atcaccccgc cgccgcattg ccgaccatcc
     2401 aacgaggagg aagtgcgtca gttcttctgg ctggccgacg tggtggtttg gcggccgctc
     2461 gagtctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     2521 acgcgtaccg gtcatcatca ccatcaccat tgagtttatc catcacactg gcggccgctc
     2581 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     2641 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2701 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2761 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2821 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2881 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2941 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3001 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3061 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3121 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3181 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3241 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3301 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3361 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3421 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3481 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3541 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3601 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3661 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3721 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3781 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3841 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3901 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3961 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4021 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4081 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4141 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4201 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4261 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4321 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4381 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4441 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4501 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4561 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4621 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4681 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4741 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4801 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4861 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4921 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4981 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5041 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5101 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5161 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5221 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5281 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5341 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5401 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5461 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5521 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5581 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5641 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5701 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5761 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5821 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5881 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5941 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6001 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6061 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6121 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6181 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6241 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6301 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6361 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6421 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6481 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6541 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6601 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6661 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6721 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6781 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6841 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6901 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6961 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7021 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     7081 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     7141 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgag
//
