LOCUS       sequence-330891-        6831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ectopic expression of hDDB2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6831)
  AUTHORS   Chen L, Bellone RR, Wang Y, Singer-Berk M, Sugasawa K, Ford JM, 
            Artandi SE
  TITLE     A novel DDB2 mutation causes defective recognition of UV-induced DNA
            damages and prevalent equine squamous cell carcinoma.
  JOURNAL   DNA Repair (Amst). 2021 Jan;97:103022. doi: 
            10.1016/j.dnarep.2020.103022. Epub 2020 Nov 12.
  PUBMED    33276309
REFERENCE   2  (bases 1 to 6831)
  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"; journalName: "DNA Repair 
            (Amst)."; date: "2021-01"; volume: "97:103022. doi"; pages: " 
            10.1016/j.dnarep.2020.103022. Epub 2020 Nov 12"
COMMENT     Sequence Label: pcDNA-hDDB2-T338M-V5
FEATURES             Location/Qualifiers
     source          1..6831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1715..1756
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1766..1783
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1806..1823)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1812..2036
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2082..2510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2169..2188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2379..2400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2519..2539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2524..2853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2704..2839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2766..2785
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2920..3714
                     /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(2974..2993)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3584..3603
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3890..4011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3927..3946)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3981..4000
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4060..4076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4060..4076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4073..4095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4084..4100
                     /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(4108..4114,4115..4132,4133..4138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4153..4174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4291..4308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4462..5050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4542..4561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5221..6012,6013..6081))
                     /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     5844..5863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6082..6186)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6261..6280)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6452..6831
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctg gctagttaag cttggtaccg
      301 agctcggatc cactagtcca gtgtggtgga attgcccttc acagtacccc ttcacacgga
      361 ggacgcgatg gctcccaaga aacgcccaga aacccagaag acctccgaga ttgtattacg
      421 ccccaggaac aagaggagca ggagtcccct ggagctggag cccgaggcca agaagctctg
      481 tgcgaagggc tccggtccta gcagaagatg tgactcagac tgcctctggg tggggctggc
      541 tggcccacag atcctgccac catgccgcag catcgtcagg accctccacc agcataagct
      601 gggcagagct tcctggccat ctgtccagca ggggctccag cagtcctttt tgcacactct
      661 ggattcttac cggatattac aaaaggctgc cccctttgac aggagggcta catccttggc
      721 gtggcaccca actcacccca gcaccgtggc tgtgggttcc aaagggggag atatcatgct
      781 ctggaatttt ggcatcaagg acaaacccac cttcatcaaa gggattggag ctggagggag
      841 catcactggg ctgaagttta accctctcaa taccaaccag ttttacgcct cctcaatgga
      901 gggaacaact aggctgcaag actttaaagg caacattcta cgagtttttg ccagctcaga
      961 caccatcaac atctggtttt gtagcctgga tgtgtctgct agtagccgaa tggtggtcac
     1021 aggagacaac gtggggaacg tgatcctgct gaacatggac ggcaaagagc tttggaatct
     1081 cagaatgcac aaaaagaaag tgacgcatgt ggccctgaac ccatgctgtg attggttcct
     1141 ggccacagcc tccgtagatc aaacagtgaa aatttgggac ctgcgccagg ttagagggaa
     1201 agccagcttc ctctactcgc tgccgcacag gcatcctgtc aacgcagctt gtttcagtcc
     1261 cgatggagcc cggctcctga ccacggacca gaagagcgag atccgagttt actctgcttc
     1321 ccagtgggac tgccccctgg gcctgatccc gcaccctcac cgtcacttcc agcacctcat
     1381 gcccatcaag gcagcctggc atcctcgcta caacctcatt gttgtgggcc gatacccaga
     1441 tcctaatttc aaaagttgta ccccttatga attgaggacg atcgacgtgt tcgatggaaa
     1501 ctcagggaag atgatgtgtc agctctatga cccagaatct tctggcatca gttcgcttaa
     1561 tgaattcaat cccatggggg acacgctggc ctctgcaatg ggttaccaca ttctcatctg
     1621 gagccaggag gaagccagga cacggaagaa gggcaattct gcagatatcc agcacagtgg
     1681 cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct
     1741 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     1801 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     1861 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     1921 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     1981 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2041 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2101 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2161 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2221 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2281 ccccaaaaaa cttgattagg gtgatggttt acgtagtggg ccatcgccct gatagacggt
     2341 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2401 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     2461 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     2521 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2581 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2641 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2701 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2761 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     2821 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     2881 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     2941 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3001 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3061 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3121 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3181 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3241 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3301 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3361 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3421 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3481 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3541 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3601 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3661 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     3721 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3781 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3841 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3901 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3961 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4021 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4081 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4141 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4201 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4261 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4321 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4381 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4621 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4681 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4801 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4861 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4921 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4981 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5041 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5101 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5161 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5221 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5281 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5341 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5401 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5461 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5521 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5581 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5641 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5701 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5761 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5821 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5881 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5941 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6001 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6061 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6121 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6181 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     6241 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     6301 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     6361 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     6421 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     6481 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     6541 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     6601 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     6661 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     6721 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     6781 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat g
//
