LOCUS       sequence-172624-        8809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8809)
  TITLE     Roussel lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8809)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCL20c-MSCV-IRES-CRE
FEATURES             Location/Qualifiers
     source          1..8809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..342
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             409..825
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     755..777
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     793..809
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    898..1447
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1042..1059)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1269..1288
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1474..2502
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     primer_bind     complement(2753..2772)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2818..2873
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2872..2891)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3234..3250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3234..3250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3247..3269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3258..3274
                     /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(3282..3288,3289..3306,3307..3312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3327..3348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3407..3602
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3453..3588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3608..3742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3645..3664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3699..3718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3810..3827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3981..4569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4061..4080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4740..5531,5532..5600))
                     /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     5363..5382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5601..5705)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5736..6115
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             6117..6374
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6421..6546
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7038..7155
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    7515..7748
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7933..7977
                     /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"
     LTR             8230..8746
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc
       61 taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt
      121 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
      181 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
      241 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
      301 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt
      361 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt
      421 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
      481 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
      541 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
      601 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
      661 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
      721 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
      781 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat tgggatccca
      841 gtgtggtggt acgtaggaat tcagatctcg agcgggatca attccgcccc ccccctaacg
      901 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
      961 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     1021 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     1081 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     1141 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     1201 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     1261 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     1321 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     1381 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     1441 gataatacca tggccaagaa gaagaggaag gtgtccaatt tactgaccgt acaccaaaat
     1501 ttgcctgcat taccggtcga tgcaacgagt gatgaggttc gcaagaacct gatggacatg
     1561 ttcagggatc gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc cgtttgccgg
     1621 tcgtgggcgg catggtgcaa gttgaataac cggaaatggt ttcccgcaga acctgaagat
     1681 gttcgcgatt atcttctata tcttcaggcg cgcggtctgg cagtaaaaac tatccagcaa
     1741 catttgggcc agctaaacat gcttcatcgt cggtccgggc tgccacgacc aagtgacagc
     1801 aatgctgttt cactggttat gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt
     1861 gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc aggttcgttc actcatggaa
     1921 aatagcgatc gctgccagga tatacgtaat ctggcatttc tggggattgc ttataacacc
     1981 ctgttacgta tagcagaaat tgccaggatc agggttaaag atatctcacg tactgacggt
     2041 gggagaatgt taatccatat tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag
     2101 aaggcactta gcctgggggt aactaaactg gtcgagcgat ggatttccgt ctctggtgta
     2161 gctgatgatc cgaataacta cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca
     2221 tctgccacca gccagctatc aactcgcgcc ctggaaggga tttttgaagc aactcatcga
     2281 ttgatttacg gcgctaagga tgactctggt cagagatacc tggcctggtc tggacacagt
     2341 gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag tttcaatacc ggagatcatg
     2401 caagctggtg gctggaccaa tgtaaatatt gtcatgaact atatccgtaa cctggatagt
     2461 gaaacagggg caatggtgcg cctgctggaa gatggcgatt aggcggccgc atcgatgccg
     2521 tatacggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     2581 aaagaaaagg ggggactgga agggctaatt cactcccaaa gaagacaaga tctgcttttt
     2641 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     2701 gggaacccac tgcttaagcc tcaataaagc ttcagctgct cgagctagca gatctttttc
     2761 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     2821 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     2881 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     2941 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     3001 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3061 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3121 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     3181 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     3241 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3301 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3361 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     3421 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3481 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3541 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3601 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3661 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3721 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     3781 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3841 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3901 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3961 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4021 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4081 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4141 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4201 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4261 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4321 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4381 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4441 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4501 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4561 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4621 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4681 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4741 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4801 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4861 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4921 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4981 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5041 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5101 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5161 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5221 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5281 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5341 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5401 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5461 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5521 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5581 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5641 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5701 ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattga ctagttatta
     5761 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     5821 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     5881 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     5941 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     6001 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6061 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgggagg
     6121 cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct
     6181 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6241 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6301 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6361 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
     6421 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     6481 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     6541 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     6601 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     6661 ttcgcagtta atactggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     6721 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     6781 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     6841 atagaggaag agcaaaacaa aagtaagaaa aaagcacagc aagcagcagg atcttcagac
     6901 ctggaaattc cctacaatcc ccaaagtcaa ggagtagtag aatctatgaa taaagaatta
     6961 aagaaaatta taggacaggt aagagatcag gctgaacatc ttaagacagc agtacaaatg
     7021 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     7081 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     7141 aaaattcaaa attttcgggt ttattacagg gacagcagaa atccactttg gaaaggacca
     7201 gcaaagctcc tctggaaagg tgaaggggca gtagtaatac aagataatag tgacataaaa
     7261 gtagtgccaa gaagaaaagc aaagatcatt agggattatg gaaaacagat ggcaggtgat
     7321 gattgtgtgg caagtagaca ggatgaggat tagaacatgg aaaagtttag taaaacacca
     7381 taaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     7441 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     7501 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     7561 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     7621 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     7681 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     7741 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     7801 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     7861 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     7921 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     7981 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     8041 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     8101 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc gagctcaagc
     8161 ttcgaacgcg ttaacgggcc cagcttcgat aaaataaaag attttattta gtctccagaa
     8221 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     8281 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     8341 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     8401 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     8461 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     8521 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     8581 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg
     8641 tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga
     8701 gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg
     8761 agatttggag acccctgccc agggaccacc gacccccccg ccgggaggt
//
