LOCUS       62056_14610        8231 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8231)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /codon_start=1
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
                     /translation="KNEQELLELDKWASL"
     promoter        1714..2214
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2688..3401
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    3447..3564
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        3682..3922
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        3939..4014
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    4088..4676
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4748..4981
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5059..5193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5220..5355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5376..5394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5404..5420)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5562..6017
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6043..6147
                     /label=AmpR promoter
     CDS             6148..7005
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7179..7767
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8055..8076
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8091..8121
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8129..8145
                     /label=lac operator
                     /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     8153..8169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8190..8208
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccttgcgcc ttttccaagg cagccctggg
     1741 tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct
     1801 gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt
     1861 gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc
     1921 gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga
     1981 cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc
     2041 tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg
     2101 gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag
     2161 cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagggggta
     2221 ccaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     2281 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     2341 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     2401 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     2461 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
     2521 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     2581 ggattcgtga attgctgccc tctggttatg tgtgggaggg cggtggaagc gccacgaact
     2641 tctctctgtt aaagcaagca ggcgacgtgg aagaaaaccc cggtcccgtg agcaagggcg
     2701 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2761 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2821 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2881 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2941 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3001 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3061 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3121 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3181 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3241 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3301 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3361 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatgaatt ctagatcttg
     3421 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     3481 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     3541 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
     3601 cgccggctcg agtgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
     3661 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
     3721 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
     3781 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
     3841 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
     3901 ttatatatct tgtggaaagg acgaaacacc gcctgagggt tttagagcta gaaatagcaa
     3961 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     4021 tctagaccca gctttcttgt acaaagttgg cattagctag cgctaaccgg tggcgcgtta
     4081 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4141 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4201 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4261 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4321 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4381 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4441 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4501 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4561 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4621 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
     4681 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     4741 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     4801 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     4861 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     4921 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     4981 agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     5041 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5101 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5161 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     5221 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5281 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5341 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg
     5401 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5461 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5521 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     5581 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5641 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5701 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5761 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5821 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5881 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5941 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6001 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6061 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6121 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6181 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6241 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6301 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6361 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6421 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6481 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6541 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6601 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6661 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6721 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6781 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6841 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6901 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6961 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7021 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7081 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7141 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7201 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7261 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7321 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7381 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7441 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7501 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7561 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7621 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7681 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7741 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7801 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7861 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7921 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     7981 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     8041 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     8101 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     8161 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     8221 gagctgcaag c
//
