LOCUS       62056_1480        9982 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 9982)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        139..1317
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1330..1725
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    1744..2332
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             2404..2637
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2715..2849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2876..3011
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3032..3050)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3218..3673
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3699..3803
                     /label=AmpR promoter
     CDS             3804..4661
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4835..5423
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5711..5732
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5747..5777
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        5846..5864
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5892..6118
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6119..6299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6346..6471
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    6964..7197
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7382..7426
                     /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        7604..7844
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        9734..9809
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    9865..9982
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
ORIGIN
        1 cgggtttatt acagggacag cagagatcca ctttggcgcc ggctcgaggg ggcccgggtg
       61 caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc
      121 ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
      181 gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag aaggtggcgc
      241 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
      301 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
      361 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc
      421 ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt
      481 cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt
      541 gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt
      601 cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct
      661 gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt
      721 atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg
      781 gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc
      841 cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg
      901 ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca
      961 gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa
     1021 aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg
     1081 ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg
     1141 gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca
     1201 gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc
     1261 attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgacgt
     1321 acggccacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     1381 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     1441 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     1501 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     1561 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     1621 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     1681 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaaac gcgttaagtc
     1741 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1801 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1861 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     1921 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     1981 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2041 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2101 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     2161 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     2221 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     2281 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
     2341 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     2401 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     2461 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     2521 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     2581 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     2641 cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     2701 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2761 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2821 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     2881 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2941 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3001 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     3061 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3121 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     3181 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     3241 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3301 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3361 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3421 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3481 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3541 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3601 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3661 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3721 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3781 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3841 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3901 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3961 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4021 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4081 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4141 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4201 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4261 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4321 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4381 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4441 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4501 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4561 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4621 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4681 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4741 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4801 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4861 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4921 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4981 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5041 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5101 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5161 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5221 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5281 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5341 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5401 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5461 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5521 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5581 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5641 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5701 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5761 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5821 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     5881 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag
     5941 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag
     6001 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac
     6061 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg
     6121 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     6181 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     6241 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     6301 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     6361 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     6421 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     6481 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     6541 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     6601 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     6661 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     6721 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     6781 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     6841 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     6901 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     6961 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     7021 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     7081 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     7141 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     7201 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     7261 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     7321 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     7381 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     7441 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     7501 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     7561 attatcgttt cagacccacc tcccaacccc gaggggaccc agagagggcc tatttcccat
     7621 gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt
     7681 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     7741 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     7801 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggagacgg
     7861 ttgtaaatga gcacacaaaa tacacatgct aaaatattat attctatgac ctttataaaa
     7921 tcaaccaaaa tcttcttttt aataacttta gtatcaataa ttagaatttt tatgttcctt
     7981 tttgcaaact tttaataaaa atgagcaaaa taaaaaaacg ctagttttag taactcgcgt
     8041 tgttttcttc acctttaata atagctactc caccacttgt tcctaagcgg tcagctcctg
     8101 cttcaatcat tttttgagca tcttcaaatg ttctaactcc accagctgct ttaactaaag
     8161 cattgtcttt aacaactgac ttcattagtt taacatcttc aaatgttgca cctgattttg
     8221 aaaatcctgt tgatgtttta acaaattcta atccagcttc aacagctatt tcacaagctt
     8281 tcatgatttc ttcttttgtt aataaacaat tttccataat acatttaaca acatgtgatc
     8341 cagctgcttt ttttacagct ttcatgtctt ctaaaactaa ttcataattt ttgtctttta
     8401 atgcaccaat atttaatacc atatcaattt ctgttgcacc atctttaatt gcttcagaaa
     8461 cttcgaatgc ttttgtagct gttgtgcatg cacctagagg aaaacctaca acatttgtta
     8521 ttcctacatt tgtgcctttt aataattctt tacaatagct tgttcaatat gaattaacac
     8581 aaactgttgc aaaatcaaat tcaattgctt catcacataa ttgtttaatt tcagctttcg
     8641 tagcatcttg ttttaataat gtgtgatcta tatatttgtt tagtttcatt ttttctccta
     8701 tatattcatt tttaatttta attctttaat aatttcgtct actttaactt tagcgttttg
     8761 aacagattca ccaacaccta taaaataaat ttttagttta ggttcagttc cacttgggcg
     8821 aacagcaaat catgacttat cttctaaata aaattttagt aagtcttgtc ctggcatatt
     8881 atacattcca tcgatgtagt cttcaacatt aacaacttta agtccagcaa tttgagttaa
     8941 gggtgttgct ctcaatgatt tcattaatgg ttcaattttt aatttctttt cttctggttt
     9001 aaaattcaag tttaaagtga aagtgtaata tgcacccatt tctttaaata aatcttctaa
     9061 atagtctact aatgttttat tttgtttttt ataaaatcaa gcagcctctg ctattaatat
     9121 agaagcttgt attccatctt tatctctagc tgagtcatca attacatatc cataactttc
     9181 ttcataagca aaaacaaaat ttaatccgtt atcttcttct ttagcaattt ctctacccat
     9241 tcatttaaat ccagttaaag tttttacaat attaactcca tatttttcat gagcgattct
     9301 atcacccaaa tcacttgtta caaaacttga atatagagcc ggattttttg gaatgctatt
     9361 taagcgtttt agatttgata attttcaatc aattaaaatt ggtcctgttt gatttccatc
     9421 taatcttaca aaatgaccat catgttttat tgccattcca aatctgtcag catctgggtc
     9481 attcataata ataatatctg catcatgttt aataccatat tcaagcggta tttttcatgc
     9541 aggatcaaat tctggatttg gatttacaac atttttaaat gtttcatctt caaatgcatg
     9601 ctcttcaacc tcaataacgt tatatcctga ttcacgtaat atttttgggg taaatttagt
     9661 tcctgttcca ttaactgcgc taaaaataat ttttaaatct tttttagctt cttgctcttt
     9721 tttgtacgtc tctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     9781 aacttgaaaa agtggcaccg agtcggtgct tttttaagct tggcgtaact agatcttgag
     9841 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
     9901 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     9961 aattacaaaa attcaaaatt tt
//
