LOCUS       sequence-330888-        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-K244E-V5
FEATURES             Location/Qualifiers
     source          1..6831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2095..2136
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2146..2163
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2186..2203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2192..2416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2462..2890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2549..2568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2759..2780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2899..2919)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2904..3233
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3084..3219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3146..3165
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3300..4094
                     /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(3354..3373)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3964..3983
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4270..4391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4307..4326)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4361..4380
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4440..4456)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4440..4456)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4453..4475)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4464..4480
                     /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(4488..4494,4495..4512,4513..4518))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4533..4554
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4671..4688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4842..5430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4922..4941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5601..6392,6393..6461))
                     /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     6224..6243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6462..6566)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6641..6660)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga attgcccttc
      721 acagtacccc ttcacacgga ggacgcgatg gctcccaaga aacgcccaga aacccagaag
      781 acctccgaga ttgtattacg ccccaggaac aagaggagca ggagtcccct ggagctggag
      841 cccgaggcca agaagctctg tgcgaagggc tccggtccta gcagaagatg tgactcagac
      901 tgcctctggg tggggctggc tggcccacag atcctgccac catgccgcag catcgtcagg
      961 accctccacc agcataagct gggcagagct tcctggccat ctgtccagca ggggctccag
     1021 cagtcctttt tgcacactct ggattcttac cggatattac aaaaggctgc cccctttgac
     1081 aggagggcta catccttggc gtggcaccca actcacccca gcaccgtggc tgtgggttcc
     1141 aaagggggag atatcatgct ctggaatttt ggcatcaagg acaaacccac cttcatcaaa
     1201 gggattggag ctggagggag catcactggg ctgaagttta accctctcaa taccaaccag
     1261 ttttacgcct cctcaatgga gggaacaact aggctgcaag actttaaagg caacattcta
     1321 cgagtttttg ccagctcaga caccatcaac atctggtttt gtagcctgga tgtgtctgct
     1381 agtagccgaa tggtggtcac aggagacaac gtggggaacg tgatcctgct gaacatggac
     1441 ggcaaagagc tttggaatct cagaatgcac aaaaaggaag tgacgcatgt ggccctgaac
     1501 ccatgctgtg attggttcct ggccacagcc tccgtagatc aaacagtgaa aatttgggac
     1561 ctgcgccagg ttagagggaa agccagcttc ctctactcgc tgccgcacag gcatcctgtc
     1621 aacgcagctt gtttcagtcc cgatggagcc cggctcctga ccacggacca gaagagcgag
     1681 atccgagttt actctgcttc ccagtgggac tgccccctgg gcctgatccc gcaccctcac
     1741 cgtcacttcc agcacctcac acccatcaag gcagcctggc atcctcgcta caacctcatt
     1801 gttgtgggcc gatacccaga tcctaatttc aaaagttgta ccccttatga attgaggacg
     1861 atcgacgtgt tcgatggaaa ctcagggaag atgatgtgtc agctctatga cccagaatct
     1921 tctggcatca gttcgcttaa tgaattcaat cccatggggg acacgctggc ctctgcaatg
     1981 ggttaccaca ttctcatctg gagccaggag gaagccagga cacggaagaa gggcaattct
     2041 gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgcggtt cgaaggtaag
     2101 cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac
     2161 cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2221 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2281 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2341 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2401 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2461 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2521 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2581 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2641 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2701 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2761 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2821 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2881 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2941 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3001 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3061 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3121 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3181 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3241 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3301 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3361 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3421 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3481 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3541 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3601 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3661 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3721 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3781 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3841 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3901 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3961 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4021 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4081 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     4141 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4201 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4261 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4321 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4381 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4441 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4501 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4561 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4621 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4681 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4741 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4801 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4861 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4921 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4981 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5041 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5101 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5161 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5221 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5281 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5341 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5401 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5461 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5521 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5581 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5641 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5701 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5761 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5821 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5881 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5941 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6001 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6061 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6121 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6181 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6241 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6301 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6361 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6421 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6481 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6541 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6601 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     6661 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     6721 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     6781 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
