LOCUS       sequence-473477-        6422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6422)
  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 6422)
  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: pcDNA3 HA c-Jun
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     388..407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(626..730)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(805..824)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        996..1375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1376..1579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1530..1550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1624..1643
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1624..1642
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1680..1706
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1731..1742)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(2736..2754)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2737..2754)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2774..2791)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2780..3004
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3050..3478
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3137..3156)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3347..3368
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3487..3507)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3492..3821
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3672..3807
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3734..3753
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3888..4682
                     /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(3942..3961)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4552..4571
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4856..4977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4893..4912)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4947..4966
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5026..5042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5026..5042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5039..5061)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5050..5066
                     /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(5074..5080,5081..5098,5099..5104))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5119..5140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5257..5274)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5428..6016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5508..5527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6187..6422,1..556,557..625))
                     /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 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
       61 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
      121 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
      181 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
      241 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
      301 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
      361 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
      421 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
      481 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
      541 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
      601 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
      661 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
      721 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
      781 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
      841 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
      901 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
      961 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     1021 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     1081 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     1141 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     1201 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     1261 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1321 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     1381 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     1441 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1501 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     1561 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
     1621 aattaatacg actcactata gggagaccca agcttggtac cgagctcgga tccaccatgt
     1681 acccatacga tgttccagat tacgctgaat tcatgactgc aaagatggaa acgaccttct
     1741 atggcgatgc cctcaacgcc tcgttcctcc cgtccgagag cggaccttat ggctacagta
     1801 accccaagat cctgaaacag agcatgaccc tgaacctggc cgacccagtg gggagcctga
     1861 agccgcacct ccgcgccaag aactcggacc tcctcacctc gcccgacgtg gggctgctca
     1921 agctggcgtc gcccgagctg gagcgcctga taatccagtc cagcaacggg cacatcacca
     1981 ccacgccgac ccccacccag ttcctgtgcc ccaagaacgt gacagatgag caggagggct
     2041 tcgccgaggg cttcgtgcgc gccctggccg aactgcacag ccagaacacg ctgcccagcg
     2101 tcacgtcggc ggcgcagccg gtcaacgggg caggcatggt ggctcccgcg gtagcctcgg
     2161 tggcaggggg cagcggcagc ggcggcttca gcgccagcct gcacagcgag ccgccggtct
     2221 acgcaaacct cagcaacttc aacccaggcg cgctgagcag cggcggcggg gcgccctcct
     2281 acggcgcggc cggcctggcc tttcccgcgc aaccccagca gcagcagcag ccgccgcacc
     2341 acctgcccca gcagatgccc gtgcagcacc cgcggctgca ggccctgaag gaggagcctc
     2401 agacagtgcc cgagatgccc ggcgagacac cgcccctgtc ccccatcgac atggagtccc
     2461 aggagcggat caaggcggag aggaagcgca tgaggaaccg catcgctgcc tccaagtgcc
     2521 gaaaaaggaa gctggagaga atcgcccggc tggaggaaaa agtgaaaacc ttgaaagctc
     2581 agaactcgga gctggcgtcc acggccaaca tgctcaggga acaggtggca cagcttaaac
     2641 agaaagtcat gaaccacgtt aacagtgggt gccaactcat gctaacgcag cagttgcaaa
     2701 cattttgact cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga
     2761 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2821 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2881 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2941 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3001 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3061 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3121 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3181 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3241 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3301 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3361 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3421 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3481 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3541 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3601 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3661 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3721 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3781 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3841 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3901 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3961 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4021 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4081 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4141 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4201 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4261 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4321 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4381 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4441 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4501 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4561 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4621 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4681 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4741 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4801 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4861 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4921 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4981 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5041 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5101 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5161 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5221 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5281 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5341 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5401 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5461 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5521 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5581 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5641 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5701 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5761 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5821 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5881 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5941 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6001 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6061 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6121 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6181 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6241 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6301 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6361 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6421 ca
//
