LOCUS       sequence-207630-        7567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shRNA for CCEPR lncRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7567)
  AUTHORS   Sharma S, Munger K
  TITLE     Expression of the cervical carcinoma expressed PCNA regulatory 
            (CCEPR) long noncoding RNA is driven by the human papillomavirus E6 
            protein and modulates cell proliferation independent of PCNA.
  JOURNAL   Virology. 2018 Feb 7;518:8-13. doi: 10.1016/j.virol.2018.01.031.
  PUBMED    29427865
REFERENCE   2  (bases 1 to 7567)
  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: "Virology.";
            date: "2018-02-7"; volume: "518:8-13. doi"; pages: " 
            10.1016/j.virol.2018.01.031"
COMMENT     Sequence Label: pLV(shRNA)-Puro-6 CCEPR Vb2
FEATURES             Location/Qualifiers
     source          1..7567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..65
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    182..203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(218..223,224..241,242..248)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    256..272
                     /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)."
     primer_bind     261..283
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     280..296
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     280..296
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     315..335
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        317..335
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        363..589
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             590..770
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    817..942
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1435..1668
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1853..1897
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2163..2280
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2287..2535
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2287..2307
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2458..2477
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    2597..2617
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        2621..3121
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3040..3058
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    3122..3146
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3153..3752
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3153..3172)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3649..3669
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(3753..3777)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3788..4376
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3841..3861)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4259..4270)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4448..4681
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4753..4874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4790..4809)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4844..4863
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4914..5049
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5069..5088)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5070..5088)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5098..5115)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5098..5114)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5107..5129)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5256..5711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5343..5362)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5553..5574
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5737..5841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5842..5910,5911..6702)
                     /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(6060..6079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6873..7461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7362..7381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
       61 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
      121 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
      181 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
      241 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
      301 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
      361 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
      421 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      481 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      541 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      601 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      661 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      721 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      781 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      841 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      901 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      961 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     1021 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     1081 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     1141 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     1201 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     1261 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     1321 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1381 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1441 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1501 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1561 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1621 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1681 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1741 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1801 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1861 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1921 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1981 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     2041 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     2101 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
     2161 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     2221 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     2281 actagtgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     2341 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     2401 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     2461 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     2521 aaaggacgaa acaccgggtg agaaatgagc ggattaccct cgagggtaat ccgctcattt
     2581 ctcacttttt gaattccaac tttgtataga aaagttgggg ttgcgccttt tccaaggcag
     2641 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc
     2701 cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc
     2761 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt
     2821 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc
     2881 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag
     2941 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg
     3001 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc
     3061 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca
     3121 ggcaagtttg tacaaaaaag caggctgcca ccatgaccga gtacaagccc acggtgcgcc
     3181 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     3241 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     3301 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     3361 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     3421 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     3481 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     3541 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     3601 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     3661 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     3721 cctggtgcat gacccgcaag cccggtgcct gaacccagct ttcttgtaca aagtggtggt
     3781 acccgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3841 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3901 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3961 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4021 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4081 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4141 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4201 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4261 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4321 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4381 gctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     4441 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     4501 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     4561 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     4621 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     4681 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     4741 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4801 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4861 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     4921 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4981 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5041 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     5101 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     5161 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5221 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     5281 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5341 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5401 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5461 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5521 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5581 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5641 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5701 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5821 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5881 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5941 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6001 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6061 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6121 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6181 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6241 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6301 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6361 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6421 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6481 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6541 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6601 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6661 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6721 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6781 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6841 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6901 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6961 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7021 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7081 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7141 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7201 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7261 gagctatgag aaagcgccac gcttcccgaa gagagaaagg cggacaggta tccggtaagc
     7321 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7381 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7441 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7501 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7561 attaccg
//
