LOCUS       Exported                5137 bp ds-DNA     circular SYN 21-NOV-2019
DEFINITION  Genomic integration vector for tetracycline-inducible expression of 
            proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5/FRT/TO
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5137)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Sep 13, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     The cloned gene must include a Kozak sequence and start codon.
FEATURES             Location/Qualifiers
     source          1..5137
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=QB0092_pRS-marker
                     /note="To sequence yeast selectable marker in pRS vectors"
     primer_bind     142..173
                     /label=OLG2812
     misc_feature    142..161
                     /label=homology_arm_Mfe1_Cut
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(301..320)
                     /label=QB1110_pSicoR
                     /note="0"
     primer_bind     566..597
                     /label=OLG2235
     misc_feature    566..590
                     /label=homology_arm_after_SNABI
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=QB0012_CMV Forward
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     776..797
                     /label=QB0976_pRLuc-MCS-F
                     /note="0"
     primer_bind     complement(796..818)
                     /label=OLG2813
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=QB0043_LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     953..981
                     /label=OLG2386
     misc_feature    953..978
                     /label=Homology after HindIII
     misc_feature    968..1077
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(968..1003)
                     /label=OLG2236
     misc_feature    983..1003
                     /label=homology_after_hindIII
     misc_feature    1057..1083
                     /label=Homology after XhoI
     primer_bind     complement(1089..1106)
                     /label=QB0008_BGH Reverse
                     /note="Bovine growth hormone terminator, reverse primer"
     polyA_signal    1095..1319
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1266..1285
                     /label=OLG2404
     primer_bind     1599..1624
                     /label=OLG2396
     protein_bind    1603..1650
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             1658..2680
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     primer_bind     1746..1765
                     /label=OLG2405
     primer_bind     1861..1889
                     /label=OLG2406
     primer_bind     complement(1885..1913)
                     /label=OLG2150
     primer_bind     2067..2086
                     /label=OLG2407
     primer_bind     2069..2093
                     /label=OLG2227
     primer_bind     2082..2107
                     /label=OLG2229
     primer_bind     2082..2107
                     /label=OLG2223
     primer_bind     2082..2106
                     /label=OLG2231
     primer_bind     2083..2107
                     /label=OLG2225
     primer_bind     2341..2373
                     /label=OLG1050
     primer_bind     complement(2464..2488)
                     /label=OLG2230
     primer_bind     2552..2576
                     /label=OLG2408
     primer_bind     2604..2632
                     /label=OLG1051
     primer_bind     complement(2614..2638)
                     /label=OLG2232
     primer_bind     complement(2614..2638)
                     /label=OLG2228
     primer_bind     complement(2614..2638)
                     /label=OLG2226
     primer_bind     complement(2614..2638)
                     /label=OLG2224
     polyA_signal    2810..2931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2830..2857)
                     /label=OLG2401
     primer_bind     complement(2847..2866)
                     /label=QB0102_SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2901..2920
                     /label=QB0017_EBV Reverse
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2980..2996)
                     /label=QB0048_M13 Reverse
                     /note="In lacZ gene"
     primer_bind     complement(2993..3016)
                     /label=QB0108_Tac promoter
                     /note="Tac promoter, forward primer"
     primer_bind     complement(2993..3015)
                     /label=QB0050_M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3211..3228)
                     /label=QB0039_L4440
                     /note="5' of MCS in L4440 vector, forward primer"
     rep_origin      complement(3382..3970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3456..3486)
                     /label=OLG2395
     primer_bind     3456..3486
                     /label=OLG2394
     primer_bind     3605..3623
                     /label=QB0766_RepO-F
                     /note="house keeping, pBR322 origin"
     primer_bind     complement(3730..3749)
                     /label=QB0767_RepO-R
                     /note="house keeping, pBR322 origin"
     CDS             complement(4141..5001)
                     /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     4764..4783
                     /label=QB0005_Amp-R
                     /note="5' end of ampicillin resistance gene, reverse 
                     primer"
     promoter        complement(5002..5106)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5115..5132
                     /label=QB0090_pQE promoter
                     /note="5' of MCS in pQE vectors, forward primer"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg
     1081 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1141 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1201 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1261 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     1321 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     1381 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1441 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1501 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1561 cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag
     1621 ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac tcaccgcgac
     1681 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     1741 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     1801 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     1861 ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta
     1921 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     1981 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca
     2041 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     2101 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     2161 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     2221 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     2281 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     2341 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     2401 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat
     2461 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     2521 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     2581 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     2641 aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac gagatttcga
     2701 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     2761 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     2821 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2881 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2941 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3001 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3061 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3121 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3181 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3241 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3301 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3361 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3421 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3481 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3541 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3601 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3661 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3721 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3781 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3841 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3901 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3961 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4021 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4081 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4141 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4201 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4261 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4321 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4381 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4441 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4501 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4561 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4621 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4681 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4741 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4801 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4861 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4921 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4981 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5101 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
