LOCUS       sequence-198791-        7103 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression vector for tetracycline-transactivator (rtTA), 
            IRES-Hygro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7103)
  AUTHORS   Khoueiry R, Sohni A, Thienpont B, Luo X, Velde JV, Bartoccetti M, 
            Boeckx B, Zwijsen A, Rao A, Lambrechts D, Koh KP
  TITLE     Lineage-specific functions of TET1 in the postimplantation mouse 
            embryo.
  JOURNAL   Nat Genet. 2017 Jul;49(7):1061-1072. doi: 10.1038/ng.3868. Epub 2017
            May 15.
  PUBMED    28504700
REFERENCE   2  (bases 1 to 7103)
  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/ng.3868"; 
            journalName: "Nat Genet"; date: "2017-07"; volume: "49"; issue: "7";
            pages: "1061-1072"
COMMENT     Sequence Label: pPB-CAG-rtTA-IRES-Hygro
FEATURES             Location/Qualifiers
     source          1..7103
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     337..359
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     421..440
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(480..1097,1098..1103,1104..1139,1140..1142,1143..1178,
                     1179..1181,1182..1217,1218..1226)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    join(1262..1836,1837..1839,1840..1848)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1429..1446)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1656..1675
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1874..2893
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             1874..2893
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     primer_bind     2998..3017
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3135..3183
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     repeat_region   3358..3420
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(3444..3462)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3483..3499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3483..3499)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3496..3518)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3507..3523
                     /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(3531..3537,3538..3555,3556..3561))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3576..3597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3714..3731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3885..4473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3965..3984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4644..5435,5436..5504))
                     /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     5267..5286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5505..5609)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5635..6090)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5772..5793)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5984..6003
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6217..6239
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6231..6248
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6232..6248
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6255..6274
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6255..6273
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   6292..6326
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        6604..6987
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
       61 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
      121 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
      181 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
      241 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
      301 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
      361 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
      421 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcctc gaggccacca
      481 tgtctagact ggacaagagc aaagtcataa acggcgctct ggaattactc aatggagtcg
      541 gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag cagcctaccc
      601 tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc gagatgctgg
      661 acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac tttctgcgga
      721 acaacgccaa gtcattccgc tgtgctctcc tctcacatcg cgacggggct aaagtgcatc
      781 tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc gcgttcctgt
      841 gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg ggccacttta
      901 cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa agagagacac
      961 ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc gaccggcagg
     1021 gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg gagaaacagc
     1081 taaagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta gacatgctcc
     1141 cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt
     1201 ttgaccttga catgctcccc gggtaagctc gagactagtt ctagagcggc cgcggatccc
     1261 gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     1321 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     1381 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     1441 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     1501 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     1561 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     1621 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     1681 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     1741 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     1801 gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaacccg taccaaagat
     1861 ggatagatcc ggaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     1921 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     1981 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     2041 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     2101 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     2161 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     2221 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     2281 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     2341 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     2401 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     2461 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     2521 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     2581 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     2641 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     2701 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     2761 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     2821 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     2881 ggcaaaggaa tagccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     2941 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3001 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3061 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3121 tcttctgagg ggatcggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     3181 ttcggatcct ttgttacttt atagaagaaa ttttgagttt ttgttttttt ttaataaata
     3241 aataaacata aataaattgt ttgttgaatt tattattagt atgtaagtgt aaatataata
     3301 aaacttaata tctattcaaa ttaataaata aacctcgata tacagaccga taaaacacat
     3361 gcgtcaattt tacgcatgat tatctttaac gtacgtcaca atatgattat ctttctaggg
     3421 ttaaagagct ccagcttttg ttccctttag tgagggttaa tttcgagctt ggcgtaatca
     3481 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3541 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3601 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3661 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3721 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3781 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3841 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3901 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3961 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4021 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4081 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4141 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4201 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4261 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4321 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4381 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4441 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4501 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4561 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4621 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4681 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4741 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4801 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4861 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4921 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4981 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5041 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5101 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5161 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5221 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5281 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5341 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5401 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5461 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5521 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5581 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     5641 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5701 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     5761 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5821 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     5881 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5941 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     6001 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     6061 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     6121 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     6181 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     6241 cggccagtga attgtaatac gactcactat agggcgaatt gggtacctta accctagaaa
     6301 gatagtctgc gtaaaattga cgcatgcatt cttgaaatat tgctctctct ttctaaatag
     6361 cgcgaatccg tcgctgtgca tttaggacat ctcagtcgcc gcttggagct cccgtgaggc
     6421 gtgcttgtca atgcggtaag tgtcactgat tttgaactat aacgaccgcg tgagtcaaaa
     6481 tgacgcatga ttatctttta cgtgactttt aagatttaac tcatacgata attatattgt
     6541 tatttcatgt tctacttacg tgataactta ttatatatat attttcttgt tatagatatc
     6601 gtcgacattg attattgact agctagttat taatagtaat caattacggg gtcattagtt
     6661 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6721 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6781 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6841 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     6901 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     6961 tacgtattag tcatcgctat taccatgatg gtcgaggtga gccccacgtt ctgcttcact
     7021 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     7081 tgtgcagcga tgggggcggg ggg
//
