LOCUS       sequence-71787-e        7429 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Retroviral Destination vector containing AttR1-AttR4 site. Use with 
            4-way LR reaction with cDNA in entry vector, marker in pBEG R2-L3 
            plasmids and shRNA in pBEG R3-L4 plasmid (module 2,3,and 4) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7429)
  AUTHORS   Geiling B, Vandal G, Posner AR, de Bruyns A, Dutchak KL, Garnett S, 
            Dankort D
  TITLE     A modular lentiviral and retroviral construction system to rapidly 
            generate vectors for gene expression and gene knockdown in vitro and
            in vivo.
  JOURNAL   PLoS One. 2013 Oct 11;8(10):e76279. doi: 
            10.1371/journal.pone.0076279.
  PUBMED    24146852
REFERENCE   2  (bases 1 to 7429)
  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: "PLoS One.";
            date: "2013-10-11"; volume: "8(10):e76279. doi"; pages: " 
            10.1371/journal.pone.0076279"
COMMENT     Sequence Label: pREG(R1-R4)   #BGV059
FEATURES             Location/Qualifiers
     source          1..7429
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        5..309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             514..689
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    752..951
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1152..1568
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1498..1520
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1536..1552
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1605..1908
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1909..2112
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2109..2133
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2220..2236
                     /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     2225..2247
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2244..2260
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2244..2260
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2291..2415
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2440..2445,2446..2463,2464..2470)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2524..3204
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(2590..2609)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3524..3829
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3708..3727
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3891..4014)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     LTR             4135..4560
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4663..4685)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4821..4841)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4826..5155
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5006..5141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5177..5196
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5277..5294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5448..6036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5528..5547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6207..6998,6999..7067))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6830..6849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7068..7172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7240..7258
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgctctg gcattatgcc
      241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      481 cgtgtacggt gggaggtcta tataagcaga gctcaataaa agagcccaca acccctcact
      541 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
      601 cttgctgttt gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga
      661 ttgactaccc acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg
      721 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt
      781 ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact
      841 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      901 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      961 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     1021 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     1081 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     1141 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1201 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1261 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1321 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1381 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1441 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1501 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1561 taggcgccgg aattgaagat ctgggggatc gatcctctag agtccgttac ataacttacg
     1621 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     1681 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1741 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     1801 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     1861 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1921 tggcagtaca tcaatgggcg tgaatagcgg tttgactcac ggggatttcc aagtctccac
     1981 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     2041 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     2101 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     2161 gacctccata gaagacaccg ggaccgatcc agcctccgcg gctcgtatgt tgtgtggaat
     2221 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccactagcag
     2281 atctgctagc acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat
     2341 caatatatta aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca
     2401 tatccagtca ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc
     2461 tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct
     2521 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     2581 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     2641 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     2701 attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac
     2761 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     2821 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     2881 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     2941 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     3001 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     3061 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     3121 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca ggcggggcgt
     3181 aatctagagg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     3241 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc
     3301 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     3361 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     3421 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     3481 gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta
     3541 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     3601 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     3661 cccccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     3721 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
     3781 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     3841 ccttatacac agccagtctg caggtcgacg atatcggcgc gccactagtc catagtgact
     3901 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     3961 tattgatatt tatatcattt tacgtttctc gttcaacttt tctatacaaa gttgcccggg
     4021 gttatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga
     4081 gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     4141 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     4201 aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagggtc
     4261 gaccctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     4321 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     4381 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     4441 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     4501 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
     4561 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
     4621 gtaagctggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4681 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4741 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     4801 cggagtgtag atccggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4861 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4921 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4981 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5041 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5101 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5161 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5221 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     5281 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5341 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5641 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5881 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6001 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6061 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     6541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     6841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7201 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7261 tttcgtcttc aagaattagc ttggccattg catacgttgt atccatatca taatatgtac
     7321 atttatattg gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat
     7381 taatagtaat caattacggg gtcattagtt catagcccat atatggagt
//
