LOCUS       sequence-257658-        8081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8081)
  AUTHORS   White EA, Sowa ME, Tan MJ, Jeudy S, Hayes SD, Santha S, Munger K, 
            Harper JW, Howley PM
  TITLE     Systematic identification of interactions between host cell proteins
            and E7 oncoproteins from diverse human papillomaviruses.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Jan 9.
  PUBMED    22232672
REFERENCE   2  (bases 1 to 8081)
  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: "Proc Natl 
            Acad Sci U S A. 2012 Jan 9."
COMMENT     Sequence Label: p6608 MSCV-IP N-HAonly CHAF1B
FEATURES             Location/Qualifiers
     source          1..8081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             326..842
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    906..1247
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1314..1730
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1660..1682
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1698..1714
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1764..1790
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1806..1830
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3508..3532)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3545..4112
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3562..4114
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3706..3723)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3933..3952
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4126..4725
                     /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(4126..4145)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4622..4642
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4801..5315
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5477..5493
                     /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(5501..5507,5508..5525,5526..5531))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5546..5567
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5684..5701)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5855..6443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5935..5954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6614..7405,7406..7474))
                     /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     7237..7256
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7475..7579)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7647..7665
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7703..7725)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7825..7844
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8038..8060
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
       61 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
      121 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
      181 agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga
      241 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
      301 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
      361 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
      421 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
      481 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
      541 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
      601 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
      661 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
      721 gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggac
      781 tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg gggtctttca
      841 tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc cccccgccgg
      901 gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg
      961 gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt atctggcgga
     1021 cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg
     1081 actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc
     1141 cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
     1201 aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca
     1261 tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagggccaga
     1321 ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc
     1381 acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc
     1441 caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg
     1501 ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg
     1561 tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta
     1621 agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga
     1681 ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc ggaattagat
     1741 ctgccaccat gctcgatgga ggatacccat acgatgttcc agattacgct gctcgaggtt
     1801 atcaaacaag tttgtacaaa aaagttggca tgaaagtcat cacttgtgaa atagcctggc
     1861 acaacaagga gcccgtgtac agcctggact tccagcatgg gacggctggg aggatccaca
     1921 gactggcgtc tgccggcgtg gacaccaatg tcaggatctg gaaggtagaa aagggaccag
     1981 atggaaaagc catcgtggaa tttttgtcca atcttgctcg tcataccaaa gccgtcaatg
     2041 ttgtgcgttt ttctccaact ggggaaattt tagcatcggg aggagatgat gctgtcatcc
     2101 tattgtggaa ggtgaatgat aacaaggagc cggagcagat cgcttttcag gatgaggacg
     2161 aggcccagct gaacaaggag aactggacgg ttgtgaagac tctgcggggc cacttagaag
     2221 atgtgtatga tatttgctgg gcaactgatg ggaatttaat ggcttctgcc tctgtggata
     2281 acacagccat catatgggat gtcagcaaag gacaaaagat atcaattttt aatgaacata
     2341 aaagttatgt ccaaggagta acctgggacc ctttgggtca atatgttgct actctgagct
     2401 gtgacagggt gctgcgagta tacagtatac agaagaagcg tgtggctttc aatgtttcga
     2461 agatgctgtc tggaataggg gctgaaggag aggcaagaag ctaccggatg tttcacgacg
     2521 acagcatgaa gtctttcttc cgtagactga gtttcactcc cgacggatct ttgcttctca
     2581 cgccagctgg atgtgtggaa tctggtgaaa atgtaatgaa taccacttat gttttctcca
     2641 ggaagaatct taaaaggccc atcgctcatc ttccatgtcc tggaaaagcc actcttgctg
     2701 ttcgctgctg tccggtctac tttgaactga ggccagtggt ggaaacaggt gtggagctga
     2761 tgagtctgcc ctaccgcctg gtgtttgctg tggcctcgga ggattccgtg cttctgtatg
     2821 acacccagca gtccttccct tttggttacg tgtctaatat acattaccac accctcagtg
     2881 acatttcatg gtccagcgat ggtgccttcc tggccatttc ttccacggac ggttactgct
     2941 catttgtgac atttgagaaa gatgaacttg gaattccttt gaaagagaag ccagttttga
     3001 acatgagaac tcctgataca gcaaagaaaa ccaagagtca gacacatcga gggtcttcgc
     3061 caggacccag accggtagag ggaacccctg ccagcagaac ccaagacccc agcagccccg
     3121 gcacgactcc ccctcaggcc agacaggccc cagccccaac agtcatcagg gaccctccct
     3181 ccatcactcc tgctgtcaaa agccccttgc cggggccttc ggaggagaag accctgcagc
     3241 ccagtagtca aaacacaaaa gcccacccat cccggagggt cactctgaac acactgcaag
     3301 cctggagcaa gacaacaccc cggagaataa acttaacacc cttaaagacg gacactccac
     3361 caagttctgt accaaccagt gtgatttcca ccccttctac agaagaaatt cagtcagaga
     3421 cgcctggaga cgctcagggc agtcccccag agctaaagcg gcccagactc gatgaaaaca
     3481 aaggaggcac ggaaagtctg gacccttgcc caactttctt gtacaaagtg gttcgataac
     3541 gaattccgcc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     3601 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     3661 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     3721 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     3781 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     3841 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     3901 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     3961 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     4021 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     4081 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatgac cgagtacaag
     4141 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4201 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4261 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4321 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4381 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4441 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4501 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4561 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4621 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4681 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgatgcat cgatgtcgac
     4741 ctgcagccaa gcttatcgat aaaataaaag attttattta gtctccagaa aaagggggga
     4801 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     4861 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
     4921 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4981 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
     5041 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     5101 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     5161 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat
     5221 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     5281 gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag
     5341 aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt
     5401 ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct
     5461 ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5521 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5581 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5641 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5701 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5761 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5821 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5881 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5941 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6001 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6061 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6121 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6181 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6241 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6301 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6361 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6421 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6481 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6541 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6601 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6661 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6721 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6781 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6841 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6901 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6961 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7021 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7081 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7141 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7201 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7261 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7321 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7381 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7441 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7501 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7561 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7621 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     7681 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     7741 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     7801 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     7861 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     7921 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     7981 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     8041 agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca t
//
