LOCUS       Exported               10082 bp ds-DNA     circular SYN 14-FEB-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10082)
  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..10082
                     /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..4136
                     /label=YAPC
     primer_bind     complement(4117..4156)
                     /label=Primer 2
     misc_feature    4155..4874
                     /label=mCherry
     CDS             4875..6368
                     /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    6472..7060
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6943..6954)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7135..7368
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        7573..7677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7678..8538
                     /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      8709..9297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9708..9729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9744..9774
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9782..9798
                     /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     9806..9822
                     /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 aggaaggtgc
     3481 gttttgccat gaaccagaga atcagtcaga gtgctccagt gaaacagcca ccacccctgg
     3541 ctccccagag cccacaggga ggcgtcatgg gtggcagcaa ctccaaccag cagcaacaga
     3601 tgcgactgca gcaactgcag atggagaagg agaggctgcg gctgaaacag caagaactgc
     3661 ttcggcaggc aatgcggaat atcaatccca gcacagcaaa ttctccaaaa tgtcaggagt
     3721 tagccctgcg tagccagtta ccaacactgg agcaggatgg tgggactcaa aatccagtgt
     3781 cttctcccgg gatgtctcag gaattgagaa caatgacgac caatagctca gatcctttcc
     3841 ttaacagtgg cacctatcac tctcgagatg agagtacaga cagtggacta agcatgagca
     3901 gctacagtgt ccctcgaacc ccagatgact tcctgaacag tgtggatgag atggatacag
     3961 gtgatactat caaccaaagc accctgccct cacagcagaa ccgtttccca gactaccttg
     4021 aagccattcc tgggacaaat gtggaccttg gaacactgga aggagatgga atgaacatag
     4081 aaggagagga gctgatgcca agtctgcagg aagctttgag ttctgacatc cttaatctcg
     4141 agtctagagg atccatggtg tctaaaggcg aggaggataa catggctata atcaaggaat
     4201 ttatgaggtt caaggtgcat atggagggat ctgtgaacgg tcacgagttc gaaatcgagg
     4261 gcgaaggcga ggggcgaccc tatgaaggga cacagacggc caaactgaag gtgaccaagg
     4321 gtggccccct gcctttcgcc tgggacatcc tgtcacccca gtttatgtac ggctctaagg
     4381 cttacgttaa gcaccctgcc gatattccgg actacttgaa actgagcttc cctgaagggt
     4441 tcaagtggga acgggtaatg aattttgagg atggcggtgt agtcacagtt acccaggaca
     4501 gctctcttca ggacggagaa tttatctata aggttaaact tcggggcact aacttcccat
     4561 cagacggccc cgtgatgcag aagaaaacta tgggctggga agccagctca gaacgcatgt
     4621 atcccgagga cggagcactg aagggagaaa tcaagcagcg actcaagctg aaggatgggg
     4681 gacattatga tgctgaggtc aaaaccacct ataaggccaa gaaacccgtt cagcttcctg
     4741 gggcatacaa cgtgaatatc aaactggata ttacatccca taacgaagac tataccatcg
     4801 tggaacagta cgagcgggcc gagggcagac atagcacagg gggcatggat gaattgtaca
     4861 aggcggccac gcgtatgaag atggacaaaa agactatagt ttggtttaga agagacctaa
     4921 ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt tttcctgtct
     4981 tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca agatggtgga
     5041 tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct gacctcactt
     5101 taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt accggtgcta
     5161 caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac cataccgtaa
     5221 aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat ctattgtatg
     5281 aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat tcttactgga
     5341 agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg cggttgatgc
     5401 caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta gggctggaga
     5461 atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggtctcca ggatggagca
     5521 atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat gcaaagaaca
     5581 gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat ttcggggaaa
     5641 taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg gcaagagata
     5701 agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt ttaagagagt
     5761 attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg ttgagtcatc
     5821 ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga caaggcagga
     5881 ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga tggatgcata
     5941 acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt ccatggaaat
     6001 ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt gacatccttg
     6061 gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc ttggacaatc
     6121 ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa tggcttcccg
     6181 agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct ttaaccgtac
     6241 tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta gacatcgaca
     6301 cagctcgtga gctactagct aaagctattt caagaacccg tgaagcacag atcatgatcg
     6361 gagcagcagc ccgggatcca ccggtcgcca ccggttcagg gagcggatcc ggctcaggga
     6421 gtgcggccgc aactcccacc tgcaacatgc gtgactgaat cgatagatcc taatcaacct
     6481 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     6541 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     6601 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     6661 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     6721 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     6781 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     6841 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     6901 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     6961 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     7021 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgagatcct ttaagaccaa
     7081 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     7141 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     7201 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     7261 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     7321 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     7381 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     7441 ccgggttaat taaggaaagg gctagatcat tcttgaagac gaaagggcct cgtgatacgc
     7501 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     7561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     7621 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     7681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     7741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     7801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     7861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     7921 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7981 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     8041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     8101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     8161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     8221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     8281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     8341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     8401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     8461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     8521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     8581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     8641 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     8701 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     8761 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     8821 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     8881 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     8941 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     9001 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     9061 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     9121 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     9181 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     9241 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     9301 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     9361 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     9421 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     9481 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcaagctc
     9541 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     9601 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctccccgtgg
     9661 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     9721 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     9781 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaatttcaca
     9841 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     9901 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc
     9961 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
    10021 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
    10081 gt
//
