LOCUS       1993_pCCLsin.PPT        8316 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 8316)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 9, 2024 from SnapGene 8.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8316
                     /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
     primer_bind     4489..4506
                     /label=lin_DNA_tgKaedeCETN1_fwd1
     misc_feature    4636..5136
                     /label=hPGK
     primer_bind     5056..5075
                     /label=pCCLsin SEQ Fwd
     primer_bind     complement(5116..5138)
                     /label=eGFPmYAP1_into_1993_A_vector_2.0
     primer_bind     5128..5187
                     /label=A-insert CETN1 transfer pl
     primer_bind     complement(5134..5157)
                     /label=A-vector CETN1 transfer pl
     primer_bind     complement(5138..5157)
                     /label=eGFPmYAP1_into_1993_A_vector
     primer_bind     5152..5171
                     /label=lin_DNA_tgKaedeCETN1_fwd2
     regulatory      5153..5161
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5158..5835
                     /codon_start=1
                     /product="Kaede green-to-red photoconvertible fluorescent 
                     protein (Ando et al., 2002)note=""direct strand 
                     translation"" 
                     translation=""MVSLIKPEMKIKLLMEGNVNGHQFVIEGDGKGHPFEGKQSMDLVV
                     KEGAPLPFAYDILTTAFHYGNRVFAKYPDHIPDYFKQSFPKGFSWERSLMFEDGGVCIA
                     TNDITLKGDTFFNKVRFDGVNFPPNGPVMQKKTLKWEASTEKMYLRDGVLTGDITMALL
                     LKGDVHYRCDFRTTYKSRQEGVKLPGYHFVDHCISILRHDKDYNEV
                     "
                     /label=Kaede
                     /translation="MVSLIKPEMKIKLLMEGNVNGHQFVIEGDGKGHPFEGKQSMDLVV
                     KEGAPLPFAYDILTTAFHYGNRVFAKYPDHIPDYFKQSFPKGFSWERSLMFEDGGVCIA
                     TNDITLKGDTFFNKVRFDGVNFPPNGPVMQKKTLKWEASTEKMYLRDGVLTGDITMALL
                     LKGDVHYRCDFRTTYKSRQEGVKLPGYHFVDHCISILRHDKDYNEVKLYEHAVAHSGLP
                     DNVK"
     primer_bind     5803..5822
                     /label=lin_DNA_tgKaedeCETN1_fwd3
     primer_bind     complement(5822..5847)
                     /label=mCETN1_into_1993_A_vector
     primer_bind     5844..5861
                     /label=whole_3143_1
     CDS             5848..6366
                     /codon_start=1
                     /product="human Centrin 1"
                     /label=Cent1
                     /translation="MASGFKKPSAASTGQKRKVAPKPELTEDQKQEVREAFDLFDVDGS
                     GTIDAKELKVAMRALGFEPRKEEMKKMISEVDREGTGKISFNDFLAVMTQKMSEKDTKE
                     EILKAFRLFDDDETGKISFKNLKRVANELGENLTDEELQEMIDEADRDGDGEVNEEEFL
                     RIMKKTSLY"
     primer_bind     complement(6341..6396)
                     /label=B-insert CETN1 transfer pl
     primer_bind     6367..6396
                     /label=B-vector CETN1 transfer pl
     primer_bind     6367..6387
                     /label=eGFPmYAP1_into_1993_B_vector
     primer_bind     6370..6392
                     /label=eGFPmYAP1_into_1993_B_vector_2.0
     misc_feature    6382..6973
                     /label=WPRE
                     /note="WPRE cds da 5983 a 6507"
     primer_bind     complement(6462..6480)
                     /label=pCCLsin SEQ Rev
     misc_feature    6970..7203
                     /label=3'LTR
                     /note="HIV-1 sin36 LTR"
     misc_feature    7110..7290
                     /label=RU5
     primer_bind     7317..7364
                     /label=mCETN1_into_1993_B_vector
     misc_feature    7362..7483
                     /label=SV40 poly(A) 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 gtgagtctga ttaaaccaga aatgaagatc aagctgctta tggaaggcaa tgtaaacggg
     5221 caccagtttg ttattgaggg agatggaaaa ggccatcctt ttgagggaaa acagagtatg
     5281 gaccttgtag tcaaagaagg cgcacctctc ccttttgcct acgatatctt gacaacagca
     5341 ttccattatg gtaacagggt ttttgctaaa tacccagacc atataccaga ctacttcaag
     5401 cagtcgtttc ccaaagggtt ttcttgggag cgaagcctga tgttcgagga cgggggcgtt
     5461 tgcatcgcta caaatgacat aacactgaaa ggagacactt tttttaacaa agttcgattt
     5521 gatggcgtaa actttccccc aaatggtcct gttatgcaga agaagactct gaaatgggag
     5581 gcatccactg agaaaatgta tttgcgtgat ggagtgttga cgggcgatat taccatggct
     5641 ctgctgctta aaggagatgt ccattaccga tgtgacttca gaactactta caaatctagg
     5701 caggagggtg tcaagttgcc aggatatcac tttgtcgatc actgcatcag catattgagg
     5761 catgacaaag actacaacga ggttaagctg tatgagcatg ctgttgccca ttctggattg
     5821 ccggacaacg tcaaggtcga cggtaccatg gcttccggct tcaagaagcc cagcgctgcc
     5881 tccaccggcc aaaagagaaa ggtggcacct aagcccgagc tcactgagga tcagaagcaa
     5941 gaagttcggg aagcatttga cctcttcgac gtggacggaa gtgggaccat cgacgcgaag
     6001 gagctgaagg tggccatgag agcgctgggc ttcgaaccca ggaaggaaga gatgaagaaa
     6061 atgatctccg aggtggacag ggaaggcacg gggaagatca gcttcaatga cttcctggcc
     6121 gtgatgacgc agaagatgtc cgagaaggac accaaagaag aaatcctgaa ggccttcagg
     6181 ctctttgatg acgatgagac cgggaagatc tcgttcaaaa acctgaagcg tgtggccaac
     6241 gagctggggg agaacctcac ggatgaggag ctgcaggaga tgatcgacga agctgatcgg
     6301 gatggggacg gcgaagtgaa cgaggaggag ttccttcgga tcatgaagaa gaccagcctt
     6361 tactgaagcg gccgcgtcga caatcaacct ctggattaca aaatttgtga aagattgact
     6421 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     6481 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     6541 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     6601 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     6661 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     6721 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     6781 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     6841 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     6901 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     6961 gcctccccgc ctggaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg
     7021 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     7081 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     7141 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     7201 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     7261 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     7321 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     7381 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     7441 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     7501 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     7561 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     7621 aggctttttt ggaggcctag gcttttgcgt cgagacgtac ccaattcgcc ctatagtgag
     7681 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     7741 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     7801 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcgac
     7861 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     7921 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7981 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     8041 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     8101 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     8161 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     8221 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     8281 gcgaatttta acaaaatatt aacgtttaca atttcc
//
