LOCUS       sequence-207631-        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-U6-CCEPR
FEATURES             Location/Qualifiers
     source          1..7567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..197
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    314..335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(350..355,356..373,374..380)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    388..404
                     /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     393..415
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     412..428
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     412..428
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     447..467
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        449..467
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        495..721
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             722..902
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    949..1074
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1567..1800
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1985..2029
                     /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    2295..2412
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2419..2667
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2419..2439
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2590..2609
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    2729..2749
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        2753..3253
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3172..3190
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    3254..3278
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3285..3884
                     /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(3285..3304)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3781..3801
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(3885..3909)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3920..4508
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3973..3993)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4391..4402)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4580..4813
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4885..5006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4922..4941)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4976..4995
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5046..5181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5201..5220)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5202..5220)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5230..5247)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5230..5246)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5239..5261)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5388..5843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5475..5494)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5685..5706
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5869..5973
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5974..6042,6043..6834)
                     /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(6192..6211)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7005..7567,1..26)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7494..7513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
       61 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
      121 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
      181 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
      241 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      301 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      361 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      421 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
      481 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
      541 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
      601 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
      661 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
      721 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
      781 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
      841 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
      901 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
      961 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     1021 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     1081 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     1141 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     1201 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     1261 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     1321 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     1381 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     1441 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     1501 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     1561 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     1621 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     1681 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     1741 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     1801 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     1861 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     1921 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     1981 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     2041 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     2101 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2161 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     2221 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg
     2281 acggtatcgc tagcttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     2341 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     2401 attcaaaatt ttactagtga gggcctattt cccatgattc cttcatattt gcatatacga
     2461 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
     2521 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     2581 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     2641 tatatcttgt ggaaaggacg aaacaccggg gcgagcatgt ttgttgttta ctcgagtaaa
     2701 caacaaacat gctcgccttt ttgaattcca actttgtata gaaaagttgg ggttgcgcct
     2761 tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa
     2821 acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg
     2881 gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg
     2941 ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac
     3001 tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc
     3061 tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt
     3121 gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc
     3181 attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga
     3241 cctctctccc caggcaagtt tgtacaaaaa agcaggctgc caccatgacc gagtacaagc
     3301 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     3361 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     3421 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     3481 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     3541 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     3601 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     3661 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     3721 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     3781 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     3841 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaacccag ctttcttgta
     3901 caaagtggtg gtacccgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3961 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     4021 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     4081 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     4141 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     4201 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     4261 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     4321 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     4381 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4441 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4501 ctccccgcat cggctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     4561 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     4621 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     4681 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     4741 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     4801 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     4861 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     4921 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4981 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     5041 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5101 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5161 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     5221 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     5281 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     5341 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     5401 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5461 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5521 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5581 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5641 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5701 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5761 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5821 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     5881 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5941 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6001 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6061 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6121 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6181 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6241 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6301 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6361 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6421 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6481 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6541 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6601 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6661 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6721 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6781 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6841 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7141 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagagagaaa ggcggacagg
     7441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7561 tgatgct
//
