LOCUS       sequence-204945-        5265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of HA-tagged RCC1 in mammalian cells (CMV 
            promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5265)
  AUTHORS   DeLuca A, Mangiacasale R, Severino A, Malquori L, Baldi A, Palena A,
            Mileo AM, Lavia P, Paggi MG
  TITLE     E1A deregulates the centrosome cycle in a Ran GTPase-dependent 
            manner.
  JOURNAL   Cancer Res. 2003 Mar 15;63(6):1430-7.
  PUBMED    12649209
REFERENCE   2  (bases 1 to 5265)
  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: "Cancer 
            Res."; date: "2003-03-15"; volume: "63(6)"; pages: "1430-7"
COMMENT     Sequence Label: pX-RCC1-HA
FEATURES             Location/Qualifiers
     source          1..5265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     583..599
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     583..599
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1897..1923
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2070..2135
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2265..2285
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2557..2691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2594..2613)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2648..2667
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2763..3218
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2850..2869)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3060..3081
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3244..3348
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3349..3417,3418..4209)
                     /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(3567..3586)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4380..4968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4869..4888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5122..5139
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagaac tagtggatcc
      601 cccgggctgc aggaattcga tatcaagctt atggcctcac ccaagcgcat agctaaaaga
      661 aggtcccccc cagcagatgc catccccaaa agcaagaagg tgaaggtctc acacaggtcc
      721 cacagcacag aacccggctt ggtgctgaca ctaggccagg gcgacgtggg ccagctgggg
      781 ctgggtgaga atgtgatgga gaggaagaag ccggccctgg tatccattcc ggaggatgtt
      841 gtgcaggctg aggctggggg catgcacacc gtgtgtctaa gcaaaagtgg ccaggtctat
      901 tccttcggct gcaatgatga gggtgccctg ggaagggaca catcagtgga gggctcggag
      961 atggtccctg ggaaagtgga gctgcaagag aaggtggtac aggtgtcagc aggagacagt
     1021 cacacagcag ccctcaccga tgatggccgt gtcttcctct ggggctcctt ccgggacaat
     1081 aacggtgtga ttggactgtt ggagcccatg aagaagagca tggtgcctgt gcaggtgcag
     1141 ctggatgtgc ctgtggtaaa ggtggcctca ggaaacgacc acttggtgat gctgacagct
     1201 gatggtgacc tctacacctt gggctgcggg gaacagggcc agctaggccg tgtgcctgag
     1261 ttatttgcca accgtggtgg ccggcaaggc ctcgaacgac tcctggtccc caagtgtgtg
     1321 atgctgaaat ccaggggaag ccggggccac gtgagattcc aggatgcctt ttgtggtgcc
     1381 tatttcacct ttgccatctc ccatgagggc cacgtgtacg gcttcggcct ctccaactac
     1441 catcagcttg gaactccggg cacagaatct tgcttcatac cccagaacct aacatccttc
     1501 aagaattcca ccaagtcctg ggtgggcttc tctggtggcc agcaccatac agtctgcatg
     1561 gattcggaag gaaaagcata cagcctgggc cgggctgagt atgggcggct gggccttgga
     1621 gagggtgctg aggagaagag catacccacc ctcatctcca ggctgcctgc tgtctcctcg
     1681 gtggcttgtg gggcctctgt ggggtatgct gtgaccaagg atggtcgtgt tttcgcctgg
     1741 ggcatgggca ccaactacca gctgggcaca gggcaggatg aggacgcctg gagccctgtg
     1801 gagatgatgg gcaaacagct ggagaaccgt gtggtcttat ctgtgtccag cgggggccag
     1861 catacagtct tattagtcaa ggacaaagaa cagagctacc catacgacgt cccagactac
     1921 gcttaggtcg acctcgaggg atcttccata cctaccagtt ctgcgcctgc aggtcgcggc
     1981 cgcgactcta gaggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca
     2041 aactacctac agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg
     2101 ttaaactact gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat
     2161 gggagcagtg gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat
     2221 ctagtgatga tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa
     2281 aggtagaaga ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt
     2341 ttagtaatag aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc
     2401 tatacaagaa aattatggaa aaatatttga tgtatagtgc cttgactaga gatcataatc
     2461 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2521 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2581 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2641 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat cctggcgtaa
     2701 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2761 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2821 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2881 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2941 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3001 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3061 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3121 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3181 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     3241 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3301 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3361 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3421 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3481 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3541 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3601 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3661 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3721 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3781 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3841 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3901 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3961 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4021 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4081 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4141 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4201 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4261 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4321 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4381 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4441 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4501 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4561 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4621 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4681 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4741 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4801 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4861 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4921 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4981 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5041 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5101 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     5161 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctagtt attaatagta
     5221 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
