LOCUS       Exported                9456 bp DNA     circular SYN 18-DEC-2024
DEFINITION  All-in-one retroviral vector with a puromycin resistance marker, for
            strong inducible expression of genes using the Tet-On(R) system.
ACCESSION   .
VERSION     .
KEYWORDS    pRetroX-TetOne-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9456)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9456)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9456)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        164..543
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    974..1331
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             1405..1812
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    1822..2193
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     polyA_signal    2298..2432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2607..2638
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        complement(2639..3006)
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     promoter        3023..3533
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3552..4295
                     /label=Tet-On(R) 3G
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of 
                     doxycycline"
     promoter        4309..4638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4647..5243
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    5264..5667
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5828..6040
                     /label=3'-LTR del in U3
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     polyA_signal    6683..6804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6827..6843)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7318..7422
                     /label=AmpR promoter
     CDS             7423..8280
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8454..9042
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9330..9351
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9366..9396
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9404..9420
                     /label=lac operator
                     /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     9428..9444
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagctagctt tgctcttagg agtttcctaa tacatcccaa actcaaatat ataaagcatt
       61 tgacttgttc tatgcgccta ggagcttggc ccattgcata cgttgtatcc atatcataat
      121 atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg attattgact
      181 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      241 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      301 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      361 tgggtggagt atttacgcta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      421 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      481 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      541 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      601 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      661 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcat ctagagcgct
      721 tctgctcccc gagctctata aaagagccca caacccctca ctcggggcgc cagtcctccg
      781 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
      841 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
      901 ggtctttcat ttgggggctc gtccgggatc gggagacccc tgcccaggga ccaccgaccc
      961 accaccggga ggtaagctgg ccagcaactt atctgtgtct gtccgattgt ctagtgtcta
     1021 tgactgattt tatgcgcctg cgtcggtact agttagctaa ctagctctgt atctggcgga
     1081 cccgtggtgg aactgacgag ttcggaacac ccggccgcaa ccctgggaga cgtcccaggg
     1141 acttcggggg ccgtttttgt ggcccgacct gagtcctaaa atcccgatcg tttaggactc
     1201 tttggtgcac cccccttaga ggagggatat gtggttctgg taggagacga gaacctaaaa
     1261 cagttcccgc ctccgtctga atttttgctt tcggtttggg accgaagccg cgccgcgcgt
     1321 cttgtctgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
     1381 gaaaatgagg gcccgggcta gcctgttacc actcccttaa gtttgacctt aggtcactgg
     1441 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     1501 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     1561 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     1621 ccagaccagg tggggtacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     1681 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     1741 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     1801 tctctaggcg cccccatatg gccatatgag atcctatatg gggcaccccc gccccttgta
     1861 aacttccctg accctgacat gacaagagtt actaacagcc cctctctcca agctcactta
     1921 caggctctct acttagtcca gcacgaagtc tggagacctc tggcggcagc ctaccaagaa
     1981 caactggacc gaccggtggt acctcaccct taccgagtcg gcgacacagt gtgggtccgc
     2041 cgacaccaga ctaagaacct agaacctcgc tggaaaggac cttacacagt cctgctgacc
     2101 acccccaccg ccctcaaagt agacggcatc gcagcttgga tacacgccgc ccacgtgaag
     2161 gctgccgacc ccgggggtgg accatcctct agatcataat cagccatacc acatttgtag
     2221 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2281 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaggcaata
     2341 gcatcacaaa tttcacaaat aaggcatttt tttcactgca ttctagtttt ggtttgtcca
     2401 aactcatcaa tgtatcttat catgtctgga tctcaaatcc ctcggaagct gcgcctgtct
     2461 taggttggag tgatacattt ttatcacttt tacccgtctt tggattaggc agtagctctg
     2521 acggccctcc tgtcttaggt tagtgaaaaa tgtcactctc ttacccgtca ttggctgtcc
     2581 agcttagctc gcaggggagg tggtctggat ccgccggcac cggtgtatac gggaattctt
     2641 tacgagggta ggaagtggta cggaaagttg gtataagaca aaagtgttgt ggaattgaag
     2701 tttactcaaa aaatcagcac tcttttatag gcgccctggt ttacataagc aaagcttata
     2761 cgttctctat cactgatagg gagtaaactg gatatacgtt ctctatcact gatagggagt
     2821 aaactgtaga tacgttctct atcactgata gggagtaaac tggtcatacg ttctctatca
     2881 ctgataggga gtaaactcct tatacgttct ctatcactga tagggagtaa agtctgcata
     2941 cgttctctat cactgatagg gagtaaactc ttcatacgtt ctctatcact gatagggagt
     3001 aaactcgagg tgataattcc acggggttgg ggttgcgcct tttccaaggc agccctgggt
     3061 ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg
     3121 ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg
     3181 tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg
     3241 cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac
     3301 agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct
     3361 cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg
     3421 cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc
     3481 gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc
     3541 atcgaattac catgtctaga ctggacaaga gcaaagtcat aaactctgct ctggaattac
     3601 tcaatggagt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg
     3661 agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa
     3721 tcgagatgct ggacaggcat catacccact cctgccccct ggaaggcgag tcatggcaag
     3781 actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg
     3841 ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc
     3901 tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg
     3961 tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg
     4021 aaagagagac acctaccacc gattctatgc ccccacttct gaaacaagca attgagctgt
     4081 tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc
     4141 tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact
     4201 tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg
     4261 ctcttgacga ttttgacctt gacatgctcc ccgggtaaac gcgcgaatgt gtgtcagtta
     4321 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4381 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4441 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4501 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4561 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4621 ggcctaggct tttgcaaaac gcgaccatga ccgagtacaa gcccacggtg cgcctcgcca
     4681 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     4741 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     4801 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     4861 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     4921 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     4981 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5041 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5101 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5161 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5221 gcatgacccg caagcccggt gcctgaacgc gtaattccct gttaatcaac ctctggatta
     5281 caaaatttgt gaaagattga ctgatattct taactatgtt gctcctttta cgctgtgtgg
     5341 atatgctgct ttaatgcctc tgtatcatgc tattgcttcc cgtacggctt tcgttttctc
     5401 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtccgtca
     5461 acgtggcgtg gtgtgctctg tgtttgctga cgcaaccccc actggctggg gcattgccac
     5521 cacctgtcaa ctcctttctg ggactttcgc tttccccctc ccgatcgcca cggcagaact
     5581 catcgccgcc tgccttgccc gctgctggac aggggctagg ttgctgggca ctgataattc
     5641 cgtggtgttg tcggggaagc tgacgtcgat tcctgcggag atatcagtgg tccaggctct
     5701 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     5761 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     5821 caagctagct tctcgcttct gttcgcgcgc ttctgctccc agacatcaac aaaagagccc
     5881 acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     5941 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     6001 tcctctgagt gattgactac ccgtcagcgg gggtctttca cgcgtatgcc gcatagttaa
     6061 gccaccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     6121 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     6181 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     6241 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     6301 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     6361 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     6421 acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg
     6481 accgaaagga gcgcacgacc ccatggctcc gaccgaagcc gacccgggcg gccccgccga
     6541 ccccgcaccc gcccccgagg cccaccgact ctagaggatc ataatcagcc ataccacatt
     6601 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     6661 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     6721 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6781 gtccaaactc atcaatgtat cttatcatgg cgcgtcgaat taattcactg gccgtcgttt
     6841 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     6901 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     6961 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     7021 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7081 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     7141 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     7201 cgtcatcacc gaaacgcgcg atgacgaaag ggcctcgtga tacgcctatt tttataggtt
     7261 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     7321 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     7381 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     7441 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     7501 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     7561 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     7621 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     7681 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7741 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7801 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7861 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7921 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7981 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     8041 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     8101 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     8161 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     8221 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     8281 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     8341 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     8401 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     8461 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     8521 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     8581 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     8641 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     8701 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8761 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8821 gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag
     8881 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8941 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     9001 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     9061 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     9121 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     9181 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     9241 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     9301 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     9361 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     9421 tttcacacag gaaacagcta tgaccatgat tacgcc
//
