LOCUS       pCGT001                 9464 bp ds-DNA     circular     09-AUG-2022
DEFINITION  .
KEYWORDS    "accession:addgene_138152_266659"
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
FEATURES             Location/Qualifiers
     rep_origin      91..679
                     /label="ori"
     primer_bind     580..599
                     /label="pBR322ori-F"
     primer_bind     833..850
                     /label="L4440"
     protein_bind    967..988
                     /label="CAP binding site"
     promoter        1003..1033
                     /label="lac promoter"
     protein_bind    1041..1057
                     /label="lac operator"
     primer_bind     1046..1068
                     /label="M13/pUC Reverse"
     primer_bind     1065..1081
                     /label="M13 Reverse"
     primer_bind     1065..1081
                     /label="M13 rev"
     primer_bind     1100..1120
                     /label="T3"
     promoter        1102..1120
                     /label="T3 promoter"
     promoter        1148..1374
                     /label="RSV promoter"
     LTR             1375..1555
                     /label="5' LTR (truncated)"
     misc_feature    1602..1727
                     /label="HIV-1 Psi"
     misc_feature    2220..2453
                     /label="RRE"
     misc_feature    2370..2389
                     /label="P0071"
     CDS             2638..2682
                     /label="gp41 peptide"
     promoter        2913..3123
                     /label="SV40 promoter"
     promoter        2935..3131
                     /label="SV40 promoter"
     rep_origin      2982..3117
                     /label="SV40 ori"
     CDS             3153..4172
                     /label="HygR"
     misc_feature    4243..4360
                     /label="cPPT/CTS"
     misc_feature    4269..4288
                     /label="P0021"
     promoter        4498..5676
                     /label="EF-1-alpha promoter"
     intron          4729..5667
                     /label="EF-1-alpha intron A"
     misc_feature    5465..5484
                     /label="P0099"
     primer          5496..5518
                     /label="ct017"
                     /note="sequence: gggggaggggttttatgcgatgg"
     primer_bind     5624..5644
                     /label="EF1a-F"
     misc_feature    5624..5644
                     /label="P0098"
     regulatory      5684..5692
                     /label="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)"
     CDS             5690..6442
                     /label="Translation 5690-6442"
                     /translation="MVPKKKRKVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGFQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNYEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYKPKKKRKV*"
     misc_feature    5693..5713
                     /label="SV40 NLS"
     primer          5714..5731
                     /label="ct001"
                     /note="sequence: gtgagcaagggcgaggag"
     CDS             5714..6418
                     /label="mOrange2"
     primer_bind     complement(5722..5731)
                     /label="Tomato-start-R"
     primer          complement(6397..6418)
                     /label="ct005"
                     /note="sequence: cttgtacagctcgtccatgccg"
     misc_feature    6464..7052
                     /label="WPRE"
     primer_bind     complement(6517..6537)
                     /label="WPRE-R"
     misc_feature    complement(6532..6551)
                     /label="P0101"
     primer          complement(6631..6654)
                     /label="ct018"
                     /note="sequence: acaacgggccacaactcctcataa"
     CDS             complement(6935..6946)
                     /label="Factor Xa site"
     LTR             7124..7357
                     /label="3' LTR (Delta-U3)"
     polyA_signal    7435..7556
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7472..7491)
                     /label="SV40pA-R"
     primer_bind     7526..7545
                     /label="EBV-rev"
     rep_origin      7596..7731
                     /label="SV40 ori"
     primer_bind     complement(7751..7770)
                     /label="T7"
     promoter        complement(7752..7770)
                     /label="T7 promoter"
     primer_bind     complement(7780..7796)
                     /label="M13 fwd"
     primer_bind     complement(7780..7797)
                     /label="M13 Forward"
     primer_bind     complement(7789..7811)
                     /label="M13/pUC Forward"
     rep_origin      7938..8393
                     /label="f1 ori"
     primer_bind     complement(8025..8044)
                     /label="F1ori-R"
     primer_bind     8235..8256
                     /label="F1ori-F"
     promoter        8419..8523
                     /label="AmpR promoter"
     CDS             8524..9384
                     /label="AmpR"
     primer_bind     complement(8742..8761)
                     /label="Amp-R"
ORIGIN
        1 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
       61 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      121 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      181 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
      241 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      301 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      361 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      421 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      481 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      541 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      601 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      661 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      721 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      781 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      841 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      901 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      961 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     1021 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     1081 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     1141 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     1201 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     1261 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     1321 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     1381 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     1441 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     1501 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1561 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1621 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1681 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1741 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1801 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1861 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1921 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1981 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     2041 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     2101 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     2161 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     2221 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     2281 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     2341 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     2401 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     2461 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     2521 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2581 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2641 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2701 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2761 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2821 tcgtttcaga cccacctccc aaccccgagg ggacccatgc attgcatctc aattagtcag
     2881 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcgtc
     2941 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3001 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3061 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3121 gcttttgcaa aaagctttct agaggtacca ccatgaaaaa gcctgaactc accgctacct
     3181 ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctccg
     3241 agggcgaaga atctcgggct ttcagcttcg atgtgggagg gcgtggatat gtcctgcggg
     3301 tgaatagctg cgccgatggt ttctacaaag atcgctatgt ttatcggcac tttgcatccg
     3361 ccgctctccc tattcccgaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt
     3421 gcatctcccg ccgtgcacag ggtgtcacct tgcaagacct gcctgaaacc gaactgcccg
     3481 ctgttctcca gcccgtcgcc gaggccatgg atgccatcgc tgccgccgat cttagccaga
     3541 ccagcgggtt cggcccattc ggacctcaag gaatcggtca atacactaca tggcgcgatt
     3601 tcatctgcgc tattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg
     3661 tcagtgcctc cgtcgcccag gctctcgatg agctgatgct ttgggccgag gactgccccg
     3721 aagtccggca cctcgtgcac gccgatttcg gctccaacaa tgtcctgacc gacaatggcc
     3781 gcataacagc cgtcattgac tggagcgagg ccatgttcgg ggattcccaa tacgaggtcg
     3841 ccaacatctt cttctggagg ccctggttgg cttgtatgga gcagcagacc cgctacttcg
     3901 agcggaggca tcccgagctt gcaggatctc ctcggctccg ggcttatatg ctccgcattg
     3961 gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca gcttgggctc
     4021 agggtcgctg cgacgcaatc gtccggtccg gagccgggac tgtcgggcgt acacaaatcg
     4081 cccgcagaag cgctgccgtc tggaccgatg gctgtgtgga agtgctcgcc gatagtggaa
     4141 acagacgccc cagcactcgt cctagggcaa aggatctgca gtaatgaggc gcgccgttaa
     4201 cgaattctag atcttgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     4261 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     4321 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     4381 cagagatcca ctttggcgcc ggctcgaggc ctgcaggtgc aaagatggat aaagttttaa
     4441 acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc
     4501 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     4561 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     4621 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     4681 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
     4741 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
     4801 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
     4861 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     4921 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     4981 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     5041 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     5101 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     5161 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
     5221 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     5281 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     5341 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     5401 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     5461 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     5521 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     5581 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     5641 gttcaaagtt tttttcttcc atttcaggtg tcgtgaggct agcgccacca tggtgcctaa
     5701 gaaaaaacgg aaagtgagca agggcgagga gaataacatg gccatcatca aggagttcat
     5761 gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     5821 gggcgagggc cgcccctacg agggctttca gaccgctaag ctgaaggtga ccaagggtgg
     5881 ccccctgccc ttcgcctggg acatcctgtc ccctcatttc acctacggct ccaaggccta
     5941 cgtgaagcac cccgccgaca tccccgacta cttcaagctg tccttccccg agggcttcaa
     6001 gtgggagcgc gtgatgaact acgaggacgg cggcgtggtg accgtgaccc aggactcctc
     6061 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     6121 cggccccgtg atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     6181 cgaggacggt gccctgaagg gcaagatcaa gatgaggctg aagctgaagg acggcggcca
     6241 ctacacctcc gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     6301 ctacatcgtc gacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     6361 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagCC
     6421 TAAGAAAAAA CGGAAAGTAT GAggatccac gcgttaagtc gacaatcaac ctctggatta
     6481 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     6541 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     6601 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     6661 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     6721 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     6781 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     6841 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     6901 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     6961 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     7021 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
     7081 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     7141 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     7201 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7261 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7321 ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat
     7381 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     7441 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7501 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7561 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     7621 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     7681 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     7741 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     7801 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     7861 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     7921 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     7981 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     8041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     8101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     8161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     8221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     8281 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     8341 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     8401 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     8461 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     8521 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     8581 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     8641 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     8701 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     8761 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     8821 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     8881 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     8941 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     9001 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     9061 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     9121 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     9181 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     9241 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     9301 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     9361 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     9421 gatttaaaac ttcattttta atttaaaagg atctaggtga agat
//