LOCUS       sequence-64549-e        8235 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 8235)
  AUTHORS   Giry-Laterriere M, Verhoeyen E, Salmon P
  TITLE     Lentiviral vectors.
  JOURNAL   Methods Mol Biol. 2011;737:183-209. doi: 
            10.1007/978-1-61779-095-9_8.
  PUBMED    21590398
REFERENCE   2  (bases 1 to 8235)
  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: "Methods Mol
            Biol."; date: "2011"; volume: "737:183-209. doi"; pages: " 
            10.1007/978-1-61779-095-9_8"
COMMENT     Sequence Label: pCLX-R4-DEST-R2
FEATURES             Location/Qualifiers
     source          1..8235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             385..642
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    689..814
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1306..1423
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    1783..2016
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2201..2245
                     /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"
     protein_bind    2522..2645
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2670..2675,2676..2693,2694..2700)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2754..3413
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2820..2839)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3755..4060
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3939..3958
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4115..4239)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4257..4845
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4310..4330)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4728..4739)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4988..5021)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5041..5221
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5259..5278)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5324..5379
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5378..5397)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5738..5754)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5738..5754)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5751..5773)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5762..5778
                     /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(5786..5792,5793..5810,5811..5816))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5831..5852
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5911..6106
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5957..6092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6112..6246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6149..6168)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6203..6222
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6314..6331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6485..7073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6565..6584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7244..8035,8036..8104))
                     /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     7867..7886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8105..8209)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggaggcg tggcctgggc gggactgggg agtggcgagc
      421 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
      481 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      541 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      601 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      661 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      721 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      781 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      841 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      901 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      961 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1021 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1081 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaaaaa
     1141 agcacagcaa gcagcaggat cttcagacct ggaaattccc tacaatcccc aaagtcaagg
     1201 agtagtagaa tctatgaata aagaattaaa gaaaattata ggacaggtaa gagatcaggc
     1261 tgaacatctt aagacagcag tacaaatggc agtattcatc cacaatttta aaagaaaagg
     1321 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     1381 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     1441 cagcagaaat ccactttgga aaggaccagc aaagctcctc tggaaaggtg aaggggcagt
     1501 agtaatacaa gataatagtg acataaaagt agtgccaaga agaaaagcaa agatcattag
     1561 ggattatgga aaacagatgg caggtgatga ttgtgtggca agtagacagg atgaggatta
     1621 gaacatggaa aagtttagta aaacaccata aggaggagat atgagggaca attggagaag
     1681 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1741 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1801 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1861 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1921 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1981 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     2041 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     2101 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     2161 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     2221 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     2281 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     2341 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     2401 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     2461 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgctagt gattatcgga
     2521 tcaactttgt atagaaaagt tgaacgagaa acgtaaaatg atataaatat caatatatta
     2581 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
     2641 ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg
     2701 tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga
     2761 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     2821 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     2881 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     2941 ttgcccgcct gatgaatgct catccggaac tccgtatggc aatgaaagac ggtgagctgg
     3001 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     3061 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     3121 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     3181 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     3241 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     3301 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     3361 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taaacgcgtg
     3421 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     3481 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     3541 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     3601 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     3661 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     3721 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     3781 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     3841 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     3901 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     3961 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     4021 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     4081 cagccagtct gcaggtcgag cagatctgct cgaccatagt gactggatat gttgtgtttt
     4141 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     4201 attttacgtt tctcgttcag ctttcttgta caaagtggtg ataatcgaat tccgataatc
     4261 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4321 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4381 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4441 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4501 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4561 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4621 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     4681 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4741 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4801 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcgg gaattcccgc
     4861 ggttcgcttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     4921 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatggg atcaattcac
     4981 catgggaata acttcgtata gcatacatta tacgaagtta tgctgctttt tgcttgtact
     5041 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5101 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5161 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5221 agcatctagt tgaatcgcca gtgtgtcgag ctagcagatc tttttccctc tgccaaaaat
     5281 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     5341 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     5401 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgccata
     5461 tgctggctgc catgaacaaa ggtggctata aagaggtcat cagtatatga aacagccccc
     5521 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     5581 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     5641 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag
     5701 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5761 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5821 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5881 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     5941 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6001 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6061 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     6121 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6181 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6241 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6301 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6361 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6421 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6481 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6541 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6601 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6661 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6721 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6781 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6841 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6901 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6961 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7021 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7081 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7141 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7201 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7261 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7321 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7381 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7441 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7501 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7561 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7621 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7681 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7741 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7801 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7861 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7921 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7981 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8041 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8101 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8161 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8221 gaaaagtgcc acctg
//
