LOCUS       sequence-267563-        6760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of Zebrafish (Zf) cytochrome 4 with a C terminal 
            FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6760)
  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 6760)
  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: pcDNA4-ZfCry4
FEATURES             Location/Qualifiers
     source          1..6760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        132..511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        512..715
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     666..686
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     760..779
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        760..778
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             853..876
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2580..2609
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2625..2642
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2665..2682)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2671..2895
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2941..3369
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3028..3047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3238..3259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3378..3398)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3383..3712
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3563..3698
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3625..3644
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3760..3807
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3826..4200
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4330..4451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4367..4386)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4421..4440
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4500..4516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4500..4516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4513..4535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4524..4540
                     /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(4548..4554,4555..4572,4573..4578))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4593..4614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4731..4748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4902..5490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4982..5001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5661..6452,6453..6521))
                     /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     6284..6303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6522..6626)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6701..6720)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
       61 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
      121 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      181 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      241 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      301 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      361 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      421 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      481 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      541 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      601 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      661 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
      721 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
      781 gacccaagct ggctagttaa gcttggtacc gagctcggat ccactagtcc agtgtggtgg
      841 aattcaatta tggactacaa ggacgacgac gacaagatga gtcaccgcac catccatctg
      901 tttcgcaaag gcctgcgcct gcatgataac cccagcctgt tgggggcgct agcgtcctcc
      961 tccgcgctct atccagtgta cgtcctggac cgtgtgttcc tgcagggggc aatgcacatg
     1021 ggtgcgttac gctggcggtt tcttctccag agtctggagg atctggacac tcgtctgcag
     1081 gcgattggat cgcgactttt tgtcctgtgc gggtcgacgg ctaacatcct gcgggagctc
     1141 gtagcacagt ggggaatcac acaaatcagt tacgacactg aagtggagcc gtattacaca
     1201 cgcatggaca aggacataca gacagtggcg caggaaaatg gactgcagac gtacacatgt
     1261 gtatcgcaca cgctgtacga cgtcaagagg atcgtgaagg ccaacggagg ctctccgccg
     1321 ctcacataca agaagttcct gcatgttttg tcagtcttgg gtgagccaga gaaaccggcg
     1381 cgggatgtga gcatcgagga cttccagagg tgtgtgactc ctgtggatgt ggatcgggtg
     1441 tatgctgtgc cttctctggc tcatctgggt ctgcaggttg aggcggaggt gctgtggccc
     1501 ggtggagaga gtcacgccct gcagcggctc gagaaacact tccagagtca gggctgggtg
     1561 gcgaacttct ccaaaccgcg caccatcccc aactctctgt tgcccagcac cactggtctg
     1621 agtccgtacc tgagtctggg ctgcctttct gttcgaacct tctaccacag actcaacagc
     1681 atctacgcac agtcgaagaa ccactctctg cccccggtgt ctctgcaggg tcaggtgttg
     1741 tggcgcgagt tcttctacac cgtcgcttcc gcaacaccaa acttcaccaa gatggaggga
     1801 aacagcatct gtctgcagat cgactggtac cacgatccag agcggctgga gaaatggaga
     1861 accgctcaga cgggttttcc ctggatcgac gccatcatga cgcagctacg gcaggagggc
     1921 tggatccatc acctggcgcg tcacgccgtg gcctgttttc tgactcgcgg agacctgtgg
     1981 atcagctggg aggagggcat gaaggtgttc gaggagttcc tgttggatgc agactacagt
     2041 gtgaacgccg ggaactggat gtggctgtcc gctagtgcgt tcttccacaa atacaccagg
     2101 atcttctgtc ctgtgcgttt cggccgtcgg actgaccctc aaggagagta tctgaggaag
     2161 tacttacctg tgctgaagaa cttcccctct cagtatattt acgagccgtg gaaagcacca
     2221 gaagacgtcc agctcagcgc cggctgcatc atcggtaaag actatccccg gccaatagtg
     2281 agccacatag aggccagcca gaggaacctg gcattgatgc gtcaggtgcg gacagagcag
     2341 cagacgaccg cagaactcac gcgagatgta gcagatgacc cgatggaggc ggggcttaag
     2401 cgtgagctcc gggaggagga ggggcttctg gaggaggcgg agtcaaagtg cacttccaag
     2461 cgtttcagtg gcagctcaga ccataagtcc cgcccctgca gctggactcc agagactcta
     2521 cagctatcag agctcagcgg agaggtcatg ggcggccgct cgagtctaga gggcccttcg
     2581 aacaaaaact catctcagaa gaggatctga atatgcatac cggtcatcat caccatcacc
     2641 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2701 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2761 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2821 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2881 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2941 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3001 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3061 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3121 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3181 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3241 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3301 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3361 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3421 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3541 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3601 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3661 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3721 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     3781 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     3841 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     3901 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     3961 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     4021 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     4081 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     4141 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     4201 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4261 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4321 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4381 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4441 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4501 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4561 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4621 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4681 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4741 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4801 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4861 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4921 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4981 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5041 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5101 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5161 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5221 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5281 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5341 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5401 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5461 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5521 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5581 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5641 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5701 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5761 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5821 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5881 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5941 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6001 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6061 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6121 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6181 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6241 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6301 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6361 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6421 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6481 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6541 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6601 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6661 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     6721 catagttaag ccagtatctg ctccctgctt gtgtgttgga
//
