LOCUS       sequence-330903-        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-R273H-V5
FEATURES             Location/Qualifiers
     source          1..6831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     702..721
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        702..720
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2168..2209
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2219..2236
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2259..2276)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2265..2489
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2535..2963
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2622..2641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2832..2853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2972..2992)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2977..3306
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3157..3292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3219..3238
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3373..4167
                     /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(3427..3446)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4037..4056
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4343..4464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4380..4399)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4434..4453
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4513..4529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4513..4529)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4526..4548)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4537..4553
                     /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(4561..4567,4568..4585,4586..4591))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4606..4627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4744..4761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4915..5503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4995..5014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5674..6465,6466..6534))
                     /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     6297..6316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6535..6639)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6714..6733)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
       61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      661 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
      721 gagacccaag ctggctagtt aagcttggta ccgagctcgg atccactagt ccagtgtggt
      781 ggaattgccc ttcacagtac cccttcacac ggaggacgcg atggctccca agaaacgccc
      841 agaaacccag aagacctccg agattgtatt acgccccagg aacaagagga gcaggagtcc
      901 cctggagctg gagcccgagg ccaagaagct ctgtgcgaag ggctccggtc ctagcagaag
      961 atgtgactca gactgcctct gggtggggct ggctggccca cagatcctgc caccatgccg
     1021 cagcatcgtc aggaccctcc accagcataa gctgggcaga gcttcctggc catctgtcca
     1081 gcaggggctc cagcagtcct ttttgcacac tctggattct taccggatat tacaaaaggc
     1141 tgcccccttt gacaggaggg ctacatcctt ggcgtggcac ccaactcacc ccagcaccgt
     1201 ggctgtgggt tccaaagggg gagatatcat gctctggaat tttggcatca aggacaaacc
     1261 caccttcatc aaagggattg gagctggagg gagcatcact gggctgaagt ttaaccctct
     1321 caataccaac cagttttacg cctcctcaat ggagggaaca actaggctgc aagactttaa
     1381 aggcaacatt ctacgagttt ttgccagctc agacaccatc aacatctggt tttgtagcct
     1441 ggatgtgtct gctagtagcc gaatggtggt cacaggagac aacgtgggga acgtgatcct
     1501 gctgaacatg gacggcaaag agctttggaa tctcagaatg cacaaaaaga aagtgacgca
     1561 tgtggccctg aacccatgct gtgattggtt cctggccaca gcctccgtag atcaaacagt
     1621 gaaaatttgg gacctgcacc aggttagagg gaaagccagc ttcctctact cgctgccgca
     1681 caggcatcct gtcaacgcag cttgtttcag tcccgatgga gcccggctcc tgaccacgga
     1741 ccagaagagc gagatccgag tttactctgc ttcccagtgg gactgccccc tgggcctgat
     1801 cccgcaccct caccgtcact tccagcacct cacacccatc aaggcagcct ggcatcctcg
     1861 ctacaacctc attgttgtgg gccgataccc agatcctaat ttcaaaagtt gtacccctta
     1921 tgaattgagg acgatcgacg tgttcgatgg aaactcaggg aagatgatgt gtcagctcta
     1981 tgacccagaa tcttctggca tcagttcgct taatgaattc aatcccatgg gggacacgct
     2041 ggcctctgca atgggttacc acattctcat ctggagccag gaggaagcca ggacacggaa
     2101 gaagggcaat tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgcg
     2161 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     2221 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     2281 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2341 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2401 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2461 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2521 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2581 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2641 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2701 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2761 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2821 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2881 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2941 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3001 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3061 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3121 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3181 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3241 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3301 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3361 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3421 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3481 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3541 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3601 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3661 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3721 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3781 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3841 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3901 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3961 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4021 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4081 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4141 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     4201 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4261 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4321 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4381 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4441 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4501 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4561 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4621 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4681 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4741 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4801 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4861 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4921 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4981 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5041 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5101 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5161 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5221 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5281 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5341 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5401 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5461 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5521 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5581 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5641 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5701 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5761 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5821 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5881 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5941 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6001 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6061 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6121 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6181 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6241 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6301 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6361 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6421 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6481 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6541 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6601 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6661 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     6721 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     6781 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga c
//
