LOCUS       sequence-140213-        7732 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 7732)
  AUTHORS   Meylan E, Dooley AL, Feldser DM, Shen L, Turk E, Ouyang C, Jacks T
  TITLE     Requirement for NF-kappaB signalling in a mouse model of lung 
            adenocarcinoma.
  JOURNAL   Nature. 2009 Nov 5;462(7269):104-7. doi: 10.1038/nature08462. Epub 
            2009 Oct 21.
  PUBMED    19847165
REFERENCE   2  (bases 1 to 7732)
  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"; doi: 
            "10.1038/nature08462"; journalName: "Nature"; date: "2009-11-5- 5"; 
            volume: "462"; issue: "7269"; pages: "104-7"
COMMENT     Sequence Label: pSicoR (Puro) p65 shRNA1
FEATURES             Location/Qualifiers
     source          1..7732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        236..615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..815
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     770..790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             833..1013
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1060..1185
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1682..1915
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2100..2144
                     /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    2442..2559
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2812..2845)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2849..2869
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     promoter        3039..3538
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3074..3097)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3471..3490
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3582..4181
                     /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(3582..3601)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4078..4098
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(4530..4563)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4619..5207
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4672..4692)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5090..5101)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5210..5226)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5211..5227)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5736..5916
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5978..6566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6058..6077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6737..7528,7529..7597))
                     /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     7360..7379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7598..7702)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct
      841 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      901 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      961 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
     1021 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
     1081 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
     1141 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
     1201 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
     1261 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
     1321 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
     1381 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
     1441 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
     1501 agtaagacca ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata
     1561 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     1621 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     1681 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     1741 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     1801 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     1861 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     1921 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     1981 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     2041 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     2101 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     2161 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     2221 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     2281 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     2341 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc
     2401 gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     2461 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     2521 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     2581 agagatccag tttggttagt accgggcccg ctctagagat ccgacgcgcc atctctaggc
     2641 ccgcgccggc cccctcgcac agacttgtgg gagaagctcg gctactcccc tgccccggtt
     2701 aatttgcata taatatttcc tagtaactat agaggcttaa tgtgcgataa aagacagata
     2761 atctgttctt tttaatacta gctacatttt acatgatagg cttggatttc tataacttcg
     2821 tatagcatac attatacgaa gttataaaca gcacaaaagg aaactcaccc taactgtaaa
     2881 gtaattgtgt gttttgagac tataaatatc ccttggagaa aagccttgtt tgaatccaga
     2941 ccaacaataa ttcaagagat tattgttggt ctggattctt ttttctcgag tactaggatc
     3001 cattaggcgg ccgaattcgt cgacctcgaa attctaccgg gtaggggagg cgcttttccc
     3061 aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg
     3121 cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg
     3181 tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca
     3241 gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat
     3301 ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct
     3361 ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag
     3421 gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac
     3481 gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac
     3541 ctgcatccat ctagatctcg atcgagcagc tgaagcttac catgaccgag tacaagccca
     3601 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     3661 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     3721 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     3781 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     3841 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     3901 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     3961 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     4021 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     4081 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     4141 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc
     4201 agcgcccgac cgaaaggagc gcacgacccc atgcatcgat gatatcataa tttaaacaag
     4261 caaaaccaaa ttaagggcca gctcattcct cccactcatg atctatagat ctatagatct
     4321 ctcgtgggat cattgttttt ctcttgattc ccactttgtg gttctaagta ctgtggtttc
     4381 caaatgtgtc agtttcatag cctgaagaac gagatcagca gcctctgttc cacatacact
     4441 tcattctcag tattgttttg ccaagttcta attccatcag aagctggtcg actctagcta
     4501 gatgcggccg caattcgtcg agggacctaa taacttcgta tagcatacat tatacgaagt
     4561 tatacatgtt taagggttcc ggttccacta ggtacaattc gatatcaagc ttatcgataa
     4621 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4681 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4741 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4801 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4861 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4921 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4981 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     5041 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5101 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5161 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     5221 acctcgatcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
     5281 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
     5341 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     5401 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
     5461 tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag
     5521 ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagcaagaga
     5581 aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg agcctgcatg
     5641 ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc ctagcatttc
     5701 atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta gaccagatct
     5761 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5821 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5881 tcagaccctt ttagtcagtg tggaaaatct ctagcagcat gtgagcaaaa ggccagcaaa
     5941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6541 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6661 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6721 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6781 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6841 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6901 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6961 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7021 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7081 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7141 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7201 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7261 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7321 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7381 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7441 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7501 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7561 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7621 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7681 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gt
//
