LOCUS       Exported                9851 bp ds-DNA     circular SYN 14-JAN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9851)
  AUTHORS   Pzx
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 26, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9851
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2224..3405
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2454..3396
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3427..3499
                     /label=Primer 1
     regulatory      3447..3456
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3459..3479
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3480..3896
                     /label=TEAD4-IDR
     primer_bind     complement(3886..3925)
                     /label=Primer 2
     CDS             4644..6137
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     misc_feature    6241..6829
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6712..6723)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6904..7137
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        7342..7446
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7447..8307
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8478..9066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9477..9498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9513..9543
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9551..9567
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9575..9591
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac gcccctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     2581 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag
     2641 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     2701 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     2761 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt
     2821 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     2881 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
     2941 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     3001 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa
     3061 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     3121 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     3181 ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     3241 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     3301 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     3361 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaagcgg ccgcaccggt
     3421 ctgcaggcgg ccgcaccggt ctgcaggcca ccatggaccc caagaagaag aggaaggtga
     3481 cagggaagac ccgcaccagg aagcaggtct ccagccacat ccaggtgctg gctcgtcgca
     3541 aagctcgcga gatccaggcc aagctaaagg accaggcagc taaggacaag gccctgcaga
     3601 gcatggctgc catgtcgtct gcacagatca tctccgccac ggccttccac agtagcatgg
     3661 ccctcgcccg gggccccggc cgcccagcag tctcagggtt ttggcaagga gctttgccag
     3721 gccaagccgg aacgtcccat gatgtgaagc ctttctctca gcaaacctat gctgtccagc
     3781 ctccgctgcc tctgccaggg tttgagtctc ctgcagggcc cgccccatcg ccctctgcgc
     3841 ccccggcacc cccatggcag ggccgcagcg tggccagctc caagctctgg atgttggagt
     3901 tctctctcga gtctagagga tccatggtgt ctaaaggcga ggaggataac atggctataa
     3961 tcaaggaatt tatgaggttc aaggtgcata tggagggatc tgtgaacggt cacgagttcg
     4021 aaatcgaggg cgaaggcgag gggcgaccct atgaagggac acagacggcc aaactgaagg
     4081 tgaccaaggg tggccccctg cctttcgcct gggacatcct gtcaccccag tttatgtacg
     4141 gctctaaggc ttacgttaag caccctgccg atattccgga ctacttgaaa ctgagcttcc
     4201 ctgaagggtt caagtgggaa cgggtaatga attttgagga tggcggtgta gtcacagtta
     4261 cccaggacag ctctcttcag gacggagaat ttatctataa ggttaaactt cggggcacta
     4321 acttcccatc agacggcccc gtgatgcaga agaaaactat gggctgggaa gccagctcag
     4381 aacgcatgta tcccgaggac ggagcactga agggagaaat caagcagcga ctcaagctga
     4441 aggatggggg acattatgat gctgaggtca aaaccaccta taaggccaag aaacccgttc
     4501 agcttcctgg ggcatacaac gtgaatatca aactggatat tacatcccat aacgaagact
     4561 ataccatcgt ggaacagtac gagcgggccg agggcagaca tagcacaggg ggcatggatg
     4621 aattgtacaa ggcggccacg cgtatgaaga tggacaaaaa gactatagtt tggtttagaa
     4681 gagacctaag gattgaggat aatcctgcat tagcagcagc tgctcacgaa ggatctgttt
     4741 ttcctgtctt catttggtgt cctgaagaag aaggacagtt ttatcctgga agagcttcaa
     4801 gatggtggat gaaacaatca cttgctcact tatctcaatc cttgaaggct cttggatctg
     4861 acctcacttt aatcaaaacc cacaacacga tttcagcgat cttggattgt atccgcgtta
     4921 ccggtgctac aaaagtcgtc tttaaccacc tctatgatcc tgtttcgtta gttcgggacc
     4981 ataccgtaaa ggagaagctg gtggaacgtg ggatctctgt gcaaagctac aatggagatc
     5041 tattgtatga accgtgggag atatactgcg aaaagggcaa accttttacg agtttcaatt
     5101 cttactggaa gaaatgctta gatatgtcga ttgaatccgt tatgcttcct cctccttggc
     5161 ggttgatgcc aataactgca gcggctgaag cgatttgggc gtgttcgatt gaagaactag
     5221 ggctggagaa tgaggccgag aaaccgagca atgcgttgtt aactagagct tggtctccag
     5281 gatggagcaa tgctgataag ttactaaatg agttcatcga gaagcagttg atagattatg
     5341 caaagaacag caagaaagtt gttgggaatt ctacttcact actttctccg tatctccatt
     5401 tcggggaaat aagcgtcaga cacgttttcc agtgtgcccg gatgaaacaa attatatggg
     5461 caagagataa gaacagtgaa ggagaagaaa gtgcagatct ttttcttagg ggaatcggtt
     5521 taagagagta ttctcggtat atatgtttca acttcccgtt tactcacgag caatcgttgt
     5581 tgagtcatct tcggtttttc ccttgggatg ctgatgttga taagttcaag gcctggagac
     5641 aaggcaggac cggttatccg ttggtggatg ccggaatgag agagctttgg gctaccggat
     5701 ggatgcataa cagaataaga gtgattgttt caagctttgc tgtgaagttt cttctccttc
     5761 catggaaatg gggaatgaag tatttctggg atacactttt ggatgctgat ttggaatgtg
     5821 acatccttgg ctggcagtat atctctggga gtatccccga tggccacgag cttgatcgct
     5881 tggacaatcc cgcgttacaa ggcgccaaat atgacccaga aggtgagtac ataaggcaat
     5941 ggcttcccga gcttgcgaga ttgccaactg aatggatcca tcatccatgg gacgctcctt
     6001 taaccgtact caaagcttct ggtgtggaac tcggaacaaa ctatgcgaaa cccattgtag
     6061 acatcgacac agctcgtgag ctactagcta aagctatttc aagaacccgt gaagcacaga
     6121 tcatgatcgg agcagcagcc cgggatccac cggtcgccac cggttcaggg agcggatccg
     6181 gctcagggag tgcggccgca actcccacct gcaacatgcg tgactgaatc gatagatcct
     6241 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     6301 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     6361 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     6421 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     6481 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     6541 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     6601 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     6661 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     6721 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     6781 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tgagatcctt
     6841 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     6901 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     6961 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     7021 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     7081 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     7141 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     7201 gtgagaggcc cgggttaatt aaggaaaggg ctagatcatt cttgaagacg aaagggcctc
     7261 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     7321 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7381 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7441 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7501 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7561 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7621 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7681 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7741 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7801 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7861 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7921 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7981 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     8041 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     8101 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     8161 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     8221 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8281 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8341 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8401 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8461 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8521 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8581 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8641 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8701 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8761 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8821 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8881 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8941 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     9001 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     9061 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     9121 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     9181 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     9241 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     9301 agcaagctca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     9361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     9421 tccccgtggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     9481 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     9541 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg
     9601 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     9661 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
     9721 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
     9781 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
     9841 aacttagtag t
//
