LOCUS       Exported                7630 bp ds-DNA     circular SYN 11-MAR-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7630)
  AUTHORS   Daniele
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 12, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7630
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      72..660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    948..969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        984..1014
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1022..1038
                     /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     1046..1062
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1083..1101
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             1357..1537
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1581..1706
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2199..2432
                     /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    2921..3038
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3061..3364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3365..3568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    3570..3588
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3591..3609
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3650..3774
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    3813..3846
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             3881..4558
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     protein_bind    4583..4616
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             4673..5389
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(5427..5551)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    5582..6170
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6053..6064)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        6226..6556
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6407..6542
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6568..6615
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6634..7008
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             7101..7334
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7406..7527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
       61 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      181 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
      241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      721 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      781 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      841 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      901 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      961 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     1021 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     1081 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
     1141 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
     1201 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
     1261 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
     1321 atattgtatt taagtgccta gctcgataca ataaacgggt ctctctggtt agaccagatc
     1381 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     1441 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     1501 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacctga
     1561 aagcgaaagg gaaaccagag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     1621 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     1681 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     1741 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     1801 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     1861 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1921 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1981 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     2041 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     2101 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     2161 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     2221 ttgggagcag caggaagcac tatgggcgca gcctcaatga cgctgacggt acaggccaga
     2281 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     2341 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     2401 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     2461 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     2521 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     2581 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     2641 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     2701 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     2761 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     2821 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     2881 acagatccat tcgattagtg aacggatctc gacggttaac ttttaaaaga aaagggggga
     2941 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     3001 aagaattaca aaaacaaatt acaaaaattc aaaattttat cgataagctt gggagttccg
     3061 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     3121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     3181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     3241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     3301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     3361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     3421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     3481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     3541 acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc tccctatcag
     3601 tgatagagat cgtcgactag tccagtgtgg tggaattctg cagatatcaa caagtttgta
     3661 caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt
     3721 gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac tatggtttaa
     3781 aggaaccaat tcagtcgact ggatccggta ccataacttc gtatagcata cattatacga
     3841 agttataatt ctgcagatat ccagcacagt ggcggccacc atggatagca ctgagaacgt
     3901 catcaagccc ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     3961 cgagatcgag ggcgagggcg agggcaagcc ctacgagggc acccagaccg ccaagctgca
     4021 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta
     4081 cggctccaag gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt
     4141 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     4201 gacccaggac tcctccctgc aggacggcac cttcatctac cacgtgaagt tcatcggcgt
     4261 gaacttcccc tccgacggcc ccgtaatgca gaagaagact ctgggctggg agccctccac
     4321 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccacaagg cgctgaagct
     4381 gaagggcggc ggccactacc tggtggagtt caagtcaatc tacatggcca agaagcccgt
     4441 gaagctgccc ggctactact acgtggactc caagctggac atcacctccc acaacgagga
     4501 ctacaccgtg gtggagcagt acgagcgcgc cgaggcccgc caccacctgt tccagtagag
     4561 ctagctgaat aaggccgctc gaataacttc gtatagcata cattatacga agttattcga
     4621 gtctagaggg cccgttagat ctctcgaggt taacgaattc gactacgcca ccatggtgag
     4681 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4741 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     4801 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4861 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     4921 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4981 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5041 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5101 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     5161 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     5221 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5281 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5341 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gtaccgaatt
     5401 cgcggccgca ctcgagatat ctagaccata gtgactggat atgttgtgtt ttacagtatt
     5461 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     5521 tttctcgttc agctttcttg tacaaagtgg ttgatatcca gcacagtggc ggccgctcga
     5581 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5641 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5701 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5761 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5821 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5881 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5941 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     6001 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6061 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6121 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattct
     6181 gcagatatcc ggttagtaat gagtttggaa ttaattctgt ggaatgtgtg tcagttaggg
     6241 tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta
     6301 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6361 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     6421 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6481 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     6541 cctaggcttt tgcaaaaagc tccccctgtt gacaattaat catcggcata gtatatcggc
     6601 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     6661 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     6721 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     6781 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     6841 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     6901 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     6961 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     7021 gatttaaatg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     7081 tttaaaagaa aaggggggac tggaagggct agctcactcc caacgaagac aagatctgct
     7141 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggctg
     7201 cctagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     7261 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     7321 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     7381 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     7441 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7501 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     7561 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     7621 gccgaggccg
//
