LOCUS       pDOXOFF-o7v1-15xGly-EGF 8454 bp ds-DNA     circular     03-FEB-2022
DEFINITION  .
COMMENT     cloning strategy: PolyGly-EGFP expression vector cloning strategy
            notebook: JY2-59, 60 samples 2a-1, 2a-3, 2a-5, 2b-4, 3a-3 derived
            from pDOXOFF-o7v1-HS-EGFP-3xHA
FEATURES             Location/Qualifiers
     rep_origin      40..175
                     /label="SV40 ori"
     promoter        175..246
                     /label="AmpR promoter"
     CDS             247..1107
                     /label="AmpR"
     rep_origin      1278..1866
                     /label="ori"
     primer          complement(1625..1643)
                     /label="QB0766 (pDOXOFF)"
                     /note="sequence: gtgtaggtcgttcgctcca"
     protein_bind    2154..2175
                     /label="CAP binding site"
     promoter        2190..2220
                     /label="lac promoter"
     protein_bind    2228..2244
                     /label="lac operator"
     primer_bind     2252..2268
                     /label="M13 rev"
     promoter        2289..2307
                     /label="T3 promoter"
     promoter        2335..2561
                     /label="RSV promoter"
     LTR             2562..2742
                     /label="5' LTR (truncated)"
     misc_feature    2789..2914
                     /label="HIV-1 Psi"
     misc_feature    3407..3640
                     /label="RRE"
     CDS             3825..3869
                     /label="gp41 peptide"
     primer          4112..4136
                     /label="QB0226 (pDOXOFF)"
                     /note="sequence: cagtgcaggggaaagaatagtagac"
     misc_feature    4276..4294
                     /label="tetO"
     misc_feature    4312..4330
                     /label="tetO"
     misc_feature    4348..4366
                     /label="tetO"
     misc_feature    4384..4402
                     /label="tetO"
     misc_feature    4420..4438
                     /label="tetO"
     misc_feature    4456..4474
                     /label="tetO"
     misc_feature    4492..4510
                     /label="tetO"
     primer          4550..4568
                     /label="RC426"
                     /note="sequence: agcagagctcgtttagtga"
     primer          4552..4572
                     /label="RC444"
                     /note="sequence: CAGAGCTCGTTTAGTGAACCG"
     primer          complement(4641..4660)
                     /label="pDOXOFF_XhoI/RPTOR_Rvs_2"
                     /note="sequence: cctggccaactgactttacg"
     primer          complement(4668..4683)
                     /label="RC339"
                     /note="sequence: ggcactaggcctggcc"
     misc_feature    4690..4695
                     /label="Kozak"
     CDS             4696..5622
                     /label="Translation 4696-5622"
                     /translation="MGLPLPGGVRQRGGGGGGGGGGGGGGGWASTVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGSGYPYDVPDYAGTGSYPYDVPDYAGSAGYPYDVPDYA*"
     primer          complement(4707..4721)
                     /label="RC443"
                     /note="sequence: acgccccctggcagc"
     primer          4713..4728
                     /label="RC444 #2"
                     /note="sequence: agggggcgtgcggcag"
     primer          4789..4806
                     /label="EGFP_Fwd"
                     /note="sequence: gtgagcaagggcgaggag"
     misc_feature    4789..5502
                     /label="EGFP"
     primer          complement(4800..4819)
                     /label="RC445"
                     /note="sequence: ccccggtgaacagctcctcg"
     primer_bind     complement(4831..4852)
                     /label="EGFP-N"
     primer_bind     complement(5092..5111)
                     /label="EXFP-R"
     primer_bind     5439..5460
                     /label="EGFP-C"
     primer          complement(5483..5502)
                     /label="EGFP_Rvs"
                     /note="sequence: cttgtacagctcgtccatgc"
     primer          5507..5526
                     /label="NLS/NES-3xHA Fwd"
                     /note="sequence: ggagcggctatccctatgac"
     misc_feature    5515..5619
                     /label="3xHA"
     primer          5620..5638
                     /label="RC340"
                     /note="sequence: TAGtgaggatccggggttg"
     promoter        5632..6142
                     /label="hPGK promoter"
     primer          complement(5648..5665)
                     /label="NLS/NES-3xHA Rvs"
                     /note="sequence: agggctgccttggaaaag"
     primer          complement(5687..5701)
                     /label="RC427"
                     /note="sequence: accacgcccagagca"
     misc_feature    6152..6553
                     /label="BlastR"
     CDS             6554..6607
                     /label="T2A"
     CDS             6608..7354
                     /label="tTA-Advanced"
     CDS             complement(7812..7823)
                     /label="Factor Xa site"
     LTR             8028..8261
                     /label="3' LTR (Delta-U3)"
     polyA_signal    8333..8454
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
       61 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
      121 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctttca
      181 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      241 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      301 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      361 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      421 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      481 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      541 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      601 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      661 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      721 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
      781 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
      841 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
      901 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
      961 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1021 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1081 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1141 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1201 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1261 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1321 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1381 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1441 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1501 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1561 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1621 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1681 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1741 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1801 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1861 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     1921 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     1981 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2041 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2101 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     2161 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     2221 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     2281 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt
     2341 cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac
     2401 aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct
     2461 tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt
     2521 gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc
     2581 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     2641 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     2701 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     2761 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     2821 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     2881 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     2941 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     3001 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     3061 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     3121 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     3181 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     3241 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     3301 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     3361 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     3421 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     3481 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     3541 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     3601 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     3661 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     3721 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     3781 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     3841 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     3901 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     3961 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     4021 acctcccaac cCCGAGGGGA CAATTCTCGA CCTCGAGACA AATGGCAGTA TTCATCCACA
     4081 ATTTTAAAAG AAAAGGGGGG ATTGGGGGGT ACAGTGCAGG GGAAAGAATA GTAGACATAA
     4141 TAGCAACAGA CATACAAACT AAAGAATTAC AAAAACAAAT TACAAAAATT CAAAATTTTC
     4201 GGGTTTATTA CAGGGACAGC AGAGATCCAC TTTGGCCGCG AATCGGGATT CGCCCTTAAG
     4261 CTAGCTCGAG TTTACTCCCT ATCAGTGATA GAGAACGTAT GAAGAGTTTA CTCCCTATCA
     4321 GTGATAGAGA ACGTATGCAG ACTTTACTCC CTATCAGTGA TAGAGAACGT ATAAGGAGTT
     4381 TACTCCCTAT CAGTGATAGA GAACGTATGA CCAGTTTACT CCCTATCAGT GATAGAGAAC
     4441 GTATCTACAG TTTACTCCCT ATCAGTGATA GAGAACGTAT ATCCAGTTTA CTCCCTATCA
     4501 GTGATAGAGA ACGTATAAGC TTTAGGCGTG TACGGTGGGC GCCTATAAAA GCAGAGCTCG
     4561 TTTAGTGAAC CGTCAGATCG CCTGGAGCAA TTCCACAACA CTTTTGTCTT ATACCAACTT
     4621 TCCGTACCAC TTCCTACCCT CGTAAAGTCA GTTGGCCAGg cctagttggc caggcctagt
     4681 gccGAATTCg ccaccATGGG GTTGCCGCTG CCAGGGGGCG TGCGGCAGCG CGGCGGCGGC
     4741 GGCGGCGGCG GCGGCGGCGG CGGCGGCGGC GGAGGCTGGG CCTCGACGgt gagcaagggc
     4801 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     4861 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     4921 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     4981 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     5041 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     5101 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     5161 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     5221 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     5281 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     5341 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     5401 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     5461 accgccgccg ggatcactct cggcatggac gagctgtaca agggagggag cggctatccc
     5521 tatgacgtgc ctgattacgc cggcacagga tcatacccct atgatgtgcc tgactacgct
     5581 ggcagcgccg gataccctta tgatgtgcct gattatgctT AGtgaggatc cggggttggg
     5641 gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg
     5701 ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag
     5761 cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc
     5821 cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc
     5881 acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc
     5941 gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga
     6001 aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg
     6061 gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg
     6121 aatcaccgac ctctctcccc agcaattcac catggttcct ttgtctcaag aagaatccac
     6181 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
     6241 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
     6301 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
     6361 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
     6421 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
     6481 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
     6541 gggccccggt gccgaaggta gaggttctct cctcacttgt ggtgatgttg aagaaaaccc
     6601 tggtccaatg tctagactgg acaagagcaa agtcataaac tctgctctgg aattactcaa
     6661 tgaagtcggt atcgaaggcc tgacgacaag gaaactcgct caaaagctgg gagttgagca
     6721 gcctaccctg tactggcacg tgaagaacaa gcgggccctg ctcgatgccc tggcaatcga
     6781 gatgctggac aggcatcata cccacttctg ccccctggaa ggcgagtcat ggcaagactt
     6841 tctgcggaac aacgccaagt cattccgctg tgctctcctc tcacatcgcg acggggctaa
     6901 agtgcatctc ggcacccgcc caacagagaa acagtacgaa accctggaaa atcagctcgc
     6961 gttcctgtgt cagcaaggct tctccctgga gaacgcactg tacgctctgt ccgccgtggg
     7021 ccactttaca ctgggctgcg tattggagga tcaggagcat caagtagcaa aagaggaaag
     7081 agagacacct accaccgatt ctatgccccc acttctgaga caagcaattg agctgttcga
     7141 ccatcaggga gccgaacctg ccttcctttt cggcctggaa ctaatcatat gtggcctgga
     7201 gaaacagcta aagtgcgaaa gcggcgggcc ggccgacgcc cttgacgatt ttgacttaga
     7261 catgctccca gccgatgccc ttgacgactt tgaccttgat atgctgcctg ctgacgctct
     7321 tgacgatttt gaccttgaca tgctccccgg gtaactaagt aaggatcgat ccaagatatc
     7381 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     7441 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     7501 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     7561 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     7621 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     7681 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     7741 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     7801 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     7861 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     7921 cctccccgcc tgtttcgcct cggcgtccgg actagaggta cctttaagac caatgactta
     7981 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     8041 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     8101 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     8161 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     8221 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     8281 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     8341 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     8401 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat ctta
//