LOCUS       sequence-473157-        6936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression in bacterial cells for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6936)
  AUTHORS   Rizzini L, Levine DC, Perelis M, Bass J, Peek CB, Pagano M
  TITLE     Cryptochromes-Mediated Inhibition of the CRL4(Cop1)-Complex Assembly
            Defines an Evolutionary Conserved Signaling Mechanism.
  JOURNAL   Curr Biol. 2019 Jun 17;29(12):1954-1962.e4. doi: 
            10.1016/j.cub.2019.04.073. Epub 2019 May 30.
  PUBMED    31155351
REFERENCE   2  (bases 1 to 6936)
  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: 
            "10.1016/j.cub.2019.04.073"; journalName: "Curr Biol"; date: 
            "2019-06-17- 17"; volume: "29"; issue: "12"; pages: "1954-1962.e4"
COMMENT     Sequence Label: pUC18 3xFLAG COP1
FEATURES             Location/Qualifiers
     source          1..6936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..119
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    236..257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(272..277,278..295,296..302)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    310..326
                     /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)."
     primer_bind     315..337
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     334..350
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     334..350
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     364..383
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        364..382
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(421..750)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      435..570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     735..755
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    779..1401
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1183..1202
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(1408..1437)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3682..3747)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3777..3980)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3806..3826)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3981..4360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4521..4538)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4521..4537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4530..4552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4678..5133)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4815..4836)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5027..5046
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5160..5264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5265..5333,5334..6125)
                     /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     complement(5483..5502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6296..6884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6785..6804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
       61 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      121 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      181 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      241 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      301 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga
      361 atttaatacg actcactata gggaattaac tttctggttt ttcagttcct cgaggagctt
      421 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
      481 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
      541 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
      601 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
      661 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
      721 tggggacttt ccacacccta actgacacac attccacagc caagctggcc gcgtacccaa
      781 ttcaacaggc atctactgag tggacccaac gcatgagagg acagtgccaa gcaagcaact
      841 caaatgtccc accggttggg catggccagg tagcctatgc tgtgtctgga cgtcctcctg
      901 ctggtatagt tattttaaaa tcagaaggac agggaaggga gcagtggttc acgcctgtaa
      961 tcccagcaat ttgggaggcc aaggtgggta gatcacctga gattaggagt tggagaccag
     1021 cctggccaat atggtgaaac cccgtctcta ccaaaaaaac aaaaattagc tgagcctggt
     1081 catgcatgcc tggaatccca acaactcggg aggctgaggc aggagaatcg cttgaaccca
     1141 ggaggcggag attgcagtga gccaagattg tgccactgca ctccagcttg gttcccaata
     1201 gaccccgcag gccctacagg ttgtcttccc aacttgcccc ttgctccata ccacccccct
     1261 ccaccccata atattataga aggacaccta gtcagacaaa atgatgcaac ttaattttat
     1321 taggacaagg ctggtgggca ctggagtggc aacttccagg gccaggagag gcactgggga
     1381 ggggtcacag ggatgccacc cgggtcacag atcctcttct gagatgagtt tttgttcgga
     1441 tcctctagag tcgactcata ccaattctag caccttaatt gtaccctgac tgttagcagc
     1501 aatcagcaca ttggactccc catctggtag tgccctccag cacacagcac taacaaattc
     1561 atttgtatca tcttcttttc ggtctttgtc gagaacactt ttgactgtat caaacttaaa
     1621 agttagcaaa gtcttagaaa gtcctttata gtacaggtag agagagttat tttcacttcc
     1681 acaagctata taatctccat tggaagccag gcctacaaag tttttttcat tgatatgacc
     1741 cttgaaggaa cgtaggcagt atggtttccc tacattccac agttttagct gactgtctgt
     1801 tgaggcagag acaatttcct caccactcac aaactttgca taagagactg ctttacggtg
     1861 tcctttgaat accatgattg gctgtttagt gttacgaaga tcatagtagt ggacacagtg
     1921 atctgcacag ccgaaagcca aatggtatct ggaagagggg ctgaatttaa cacagcacac
     1981 attagccttt gcctcaatgc ttgccactga gttgtctaga ttggtagacc acagcttcac
     2041 ttttgcatca tcagaacctg aagccaagag tttaggatcc atcaaattaa agtcaacact
     2101 ccaacacctc ttctcatgct cctgatagac ctttgacctc tgtcctgtga atccatccca
     2161 taaaataaca gtgccttcat aatcactgct agctaacagg ttcttatggt aactactcca
     2221 actgatacag ctgattttcg aattgcaggt catttcattc tcagggtaat gaatatccac
     2281 tgcatcctgg atgacagtgt catattcata gactttaatc ttctttgtaa ctccagcaat
     2341 cgcaaaatag tcacaatccc ggtcaaattc aatactagag actatactgg aaccattata
     2401 gagatcacta gcatatgaca atgtggctaa aggtcgtact gaattatatc gagtaaactt
     2461 ggacaagcat tcctgaaatt catccaactg gcttgcagtt cgactgtcat ctgagatacg
     2521 agacatcctt gtagaaaagt aacactgctc caagtcttca aaatgagcag taagtcgttt
     2581 tcgtcttgat gctaacgtgc tattatacca aggctgtttc tttgtctgag aactgccact
     2641 gaaacctgga ggttggctgt attctgtgga atcaataata ctactgtgtg atggagaagg
     2701 agcttcaaat tgaggcactg tgctatcctc actgacagga gagtataagc cactcatttc
     2761 ttccactctc ttaatatcct cttccaaaac acttagctcc ttctggatct gttccagttg
     2821 ctctctctta tttcttcttg caaccttgag gaattccata agaatctgta gttgggctgc
     2881 atgtgattct gcttccagtt gtttcttctt ctgcactagt aactccaaca taagattgac
     2941 attggccaaa tcaaggttat cttggtcagt tcccaaccaa tcttgaaata tctgccacct
     3001 gtggccattg gtgctactca ctgagtggtc caatttgaac ctcttttcct caaatctttg
     3061 cttctgttta agaatgagtt cattcaccaa gaaattagga tacagatggt caatattgtc
     3121 cacaacatag ttacacttgg gacatctatt attgtcctcc aaactctgat gaatacactt
     3181 gtagcaaaag ctgtggccac attttgtcat gtatgcttct tcaatcatat caaagcagat
     3241 ggggcatacg aagtcgttgc ttttgtcctc gtaggagttg atgagcccgt tgcagagggg
     3301 ggcgaggaga ggtcgcttcc tgctgccgct gcctaggctg gagctgctgc ctcctacgcc
     3361 ggcgctgggc ctggccgcgc agctgtgccg ggacaggccc gtggacaccg ccccgccgcc
     3421 gctacccgat acggcgggcg ccaccaacac aggccgcacc gggcccccga ggccgcccga
     3481 gccggcggcc tgggccaccc cgccggacac cagcgctgcc gccgaaaccg ccacggaagg
     3541 cggcgacggg gaagaggata aagacgagga ggcggaagtc accgaggagg ccgccgagga
     3601 cccggggctt gtcccagcgg agcccgaccc ggcctggcgg ctaccagaca ttagatctat
     3661 cgatgaattc gcggccgcaa gcttgtcatc gtcatccttg taatcgatgt catgatcttt
     3721 ataatcaccg tcatggtctt tgtagtccat ggttaattct gacggttcac taaacgagct
     3781 ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa cggggcgggg
     3841 ttattacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg
     3901 tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattggtg
     3961 tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa
     4021 gtaggaaagt cccgtaaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     4081 ttgacgtcaa tagggggcgg acttggcata tgatacactt gatgtactgc caagtgggca
     4141 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     4201 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     4261 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     4321 gtaattgatt actattaata actagtcaat aatcaatgtc aacatggcgg tcatattgga
     4381 catgagccaa tataaatgta catattatga tatagataca acgtatgcaa tggccaatag
     4441 ccaatattga tttatgctat ataaccaatg actaatatgg ctaattgcca atattgattc
     4501 aatgtataga tcagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     4561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4621 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa
     4681 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4741 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4801 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4861 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4921 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4981 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5041 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5101 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     5161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6121 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6421 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6721 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6841 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6901 cctttttacg gttcctggcc ttttgctggc cttttg
//
