LOCUS       1996_pCCLsin.PPT        9945 bp    DNA     circular SYN 12-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9945)
  AUTHORS   Department of Biology
  TITLE     Direct Submission
  JOURNAL   Exported Dec 9, 2024 from SnapGene 8.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9945
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    132..992
                     /label=AmpR
     misc_feature    2338..2876
                     /label=CMV IE
     misc_feature    2891..3071
                     /label=RU5
     misc_feature    2930..3584
                     /label=5'LTR
     misc_feature    3120..3257
                     /label=psi
                     /note="psi"
     misc_feature    3177..3180
                     /label=5' splice signal(donor)
                     /note="5' splice signal(donor)"
     misc_feature    3224..3588
                     /label=gag gene mutated
                     /note="gag gene mutated"
     misc_feature    3744..3948
                     /label=RRE
     misc_feature    4340..4342
                     /label=3' splicing site(acceptor)
                     /note="3' splicing site(acceptor)"
     misc_feature    4457..4570
                     /label=cPPT
     misc_feature    4636..5136
                     /label=hPGK
                     /note="Sequence ok except for point mutation A to G at bp 
                     no. 333"
     primer_bind     5056..5075
                     /label=pCCLsin SEQ Fwd
     primer_bind     5116..5179
                     /label=eGFPmYAP1_into_270_A_insert_3.0
     primer_bind     5116..5178
                     /label=mVenus_hCEP83_into_270_A_insert
     primer_bind     5116..5164
                     /label=eGFPmYAP1_into_1993_A_insert_2.0
     primer_bind     complement(5134..5157)
                     /label=A-vector CETN1 transfer pl
     primer_bind     5138..5177
                     /label=eGFPmYAP1_into_1993_A_insert
     primer_bind     5138..5173
                     /label=eGFPmCEP83_into_270_A_insert
     regulatory      5153..5161
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5158..5160,5161..5163,5164..5874)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     5158..5183
                     /label=eGFPmYAP1_into_TwistKaede_A_vector
     primer_bind     complement(5203..5224)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5464..5483)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5780..5799
                     /label=17843 pEGFP C3 mYAP1 SEQ FWD2
     primer_bind     5811..5832
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5858..5877
                     /label=mVenus_hCEP83_Fwd1
     primer_bind     complement(5896..5908)
                     /label=A-vector YAP1 Gibbson
     misc_feature    5920..7995
                     /label=Mus musculus Cep83
     primer_bind     6631..6650
                     /label=mVenus_hCEP83_Fwd2
     primer_bind     complement(7967..8023)
                     /label=eGFPmCEP83_into_270_B_insert
     primer_bind     7996..8025
                     /label=B-vector CETN1 transfer pl
     misc_feature    8011..8602
                     /label=WPRE
                     /note="WPRE cds da 5983 a 6507"
     primer_bind     complement(8091..8109)
                     /label=pCCLsin SEQ Rev
     misc_feature    8599..8832
                     /label=3'LTR
                     /note="HIV-1 sin36 LTR"
     misc_feature    8739..8919
                     /label=RU5
     polyA_signal    8991..9112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttggc
     2221 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat
     2281 taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     2341 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     2401 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     2461 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     2521 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     2581 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     2641 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     2701 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     2761 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     2821 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     2881 tagtgaaccg gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     2941 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     3001 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     3061 aatctctagc agtggcgccc gaacagggac ctgaaagcga aagggaaacc agagctctct
     3121 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     3181 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     3241 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     3301 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     3361 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     3421 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     3481 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     3541 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
     3601 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     3661 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     3721 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     3781 cgcagcctca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     3841 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     3901 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     3961 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     4021 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     4081 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     4141 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     4201 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     4261 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     4321 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     4381 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     4441 atctcgacgg tatcggttaa cttttaaaag aaaagggggg attggggggt acagtgcagg
     4501 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     4561 tacaaaaatt caaaatttta tcgatcacga gactagcctc gagaagcttg atatcgaatt
     4621 cccacggggt tggggttgcg ccttttccaa ggcagccctg ggtttgcgca gggacgcggc
     4681 tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc
     4741 acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc cccggcgacg
     4801 cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga
     4861 caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca gggagcaatg
     4921 gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcggg gcgcgccgag
     4981 agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg
     5041 ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc
     5101 tccctcgttg accgaatcac cgacctctct ccccaggggg atccaccggt cgccaccatg
     5161 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     5221 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     5281 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     5341 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     5401 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     5461 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     5521 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     5581 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     5641 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     5701 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     5761 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     5821 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga
     5881 ctcagatctc gagctcaagc ttcgaattct gcagtcgacg acacatttcc tagcctattt
     5941 ccacctggag gagatagtag gctgaatcct gagcctgagt tccaaaatat gttaattgat
     6001 gaaagggtac gctgtgaaca tcataaacat aattatcagg ctctgaaaat tgaacacaaa
     6061 aggttgcagg aagaatatgt aaaatcacaa aatgaactta aacgtgtatt aattgaaaag
     6121 caagcaagcc aggaaaaatt ccaactgctc cttgaagact taaggggaga attagtagag
     6181 aaagctagag acatagaaaa aatgaaactg caggtactaa caccacaaaa attggaattg
     6241 gtaaaagccc aactacaaca agaattagaa gctccaatgc gagaacgttt tcggactctt
     6301 gatgaagaag tggaaaggta cagagctgag tataacaagc tgcgctacga gtatacattt
     6361 ctcaagtcag agtttgaaca ccagaaagaa gagtttactc gggtttcaga agaagagaaa
     6421 atgaaataca agtcagaggt tgcacgactg gagaaggaca aagaggagct acataaccag
     6481 ctgcttagtg ttgatcccac gagagacagc aaacgaatgg agcaacttgt tcgagaaaaa
     6541 acccatttgc ttcagaaatt gaaaagttta gaggctgaag tagcagaatt aagggctgag
     6601 aaagaaaatt ctggtgctca ggtagaaaat gtccaaagaa tacaggtgag gcagttggct
     6661 gagatgcagg ctacactcag atccttggag gctgaaaagc agtcagctaa actacaagct
     6721 gagcgtttag aaaaagaact acaatcaagc aatgaacaga atacctgctt aatcagcaaa
     6781 ctgcatagag ctgaccgaga aatcagcaca ctggccagtg aagtgaaaga gcttaaacat
     6841 gcaaacaaac tagaaataac tgacatcaaa ctggaggcag caagagctaa gagtgagctc
     6901 gaaagagaaa ggaataagat ccaaagtgaa cttgatggat tgcagtcaga caatgaaatt
     6961 ctcaagtcaa ctgttgaaca ccacaaagcg ctcttagtgg aaaaagatcg tgaattaata
     7021 cgtaaagtac aagctgccaa ggaagaaggt taccagaaac tcatggtctt acaagatgaa
     7081 aagctagaac ttgagaacag attatcagac ttagagaaga tgaaggtgga gcgtgatgtc
     7141 tggaggcagt cggaaaagga gcagtgtgag gagaagctgc gggcctccca gatggcagag
     7201 gaggcagccc gcagggagct gcagagcact aggttaaaac ttcaacaaca aattgtgaat
     7261 accgaaaaag cagaaaaaga aaaacttgaa aattctgaac tgaaacagca aatcagtcat
     7321 ttgcagatcc aagtgacctc actgacacag tcggagaacg acttactgaa ttcaaaccac
     7381 atgctgaagg acatggtgga gagactaaaa caagaatgcc gaaatttaag gagccaagct
     7441 gagaaagcac aactagacgt tgaaaagaca ctggaggaaa aacagatcca gtggttggaa
     7501 gaaaagcaca agctccatga gcgaatcaca gacagagaag aaaagtataa ccaagccaag
     7561 gaaaagctac agcgcgcagc gactgctcag aaaaagagaa aatctcttca tgagaacaaa
     7621 ttgaagagac tacaggagaa agtagaagtc ttggaggcaa agaaagaaga actagaaaca
     7681 gaaaaccagg tgctaaatag acaaaatgtt ccatttgaag aatatacaag gcttcaaaaa
     7741 cgactaaagg acatacagag aagacataat gaatttcgaa gtctaatttt ggttcctaac
     7801 atgcctccaa cagcatcgat tagtcctgct aacttccagt ccgcagtgac agttcctgga
     7861 gcggaactct cttttccccc tcatctgcag gaggaacaac atcaaagaga gctctctctc
     7921 cttcgcaaaa gactggaaga actggagact acacaaagaa aacaactgga ggagcttggg
     7981 tctcctgggg agtgaagcgg ccgcgtcgac aatcaacctc tggattacaa aatttgtgaa
     8041 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     8101 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     8161 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     8221 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     8281 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     8341 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     8401 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     8461 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     8521 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     8581 ctttgggccg cctccccgcc tggaattcga gctcggtacc tttaagacca atgacttaca
     8641 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     8701 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     8761 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     8821 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     8881 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     8941 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     9001 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     9061 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     9121 ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     9181 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     9241 gtagtgagga ggcttttttg gaggcctagg cttttgcgtc gagacgtacc caattcgccc
     9301 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     9361 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     9421 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     9481 tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     9541 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     9601 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     9661 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     9721 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     9781 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     9841 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     9901 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcc
//
