LOCUS       V025529                 9028 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025529
VERSION     V025529
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2284..2991
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             3028..4125
                     /label="Sms(NM_009214)"
                     /note="Sms(NM_009214)"
                     /gene="Sms"
     CDS             4126..4152
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4193..4404
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4417..4685
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4718..5116
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5123..5711
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5785..6018
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6090..6211
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6230..6365
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6403..6419)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6427..6443
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6451..6481)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6496..6517
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6805..7393)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7564..8424)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8425..8529)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9003..9019
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagatgcc
     2281 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     2341 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2401 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2461 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2521 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2581 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2641 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2701 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2761 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2821 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2881 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2941 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gggaggtgga
     3001 ggtagtggag gaggcggatc gtctagaatg gcagcagcaa gacacagcac gctcgacttc
     3061 aagctcggcg ccaaagctga tggtgaggcc attctgaaag gccttcagtc catttttcag
     3121 gagcagggga tgacagagtc agtgcacacc tggcaggacc atggatattt agcaacttac
     3181 acgaacaaga atggcagctt tgccaatttg cgaatttatc cacatggatt ggtgttgctg
     3241 gaccttcaga gttacgacag tgatgtgcaa ggcaaacaag aaactgacag ccttttgaac
     3301 aaaatagaag aaaaaatgaa agaattgagt caagacagta ctgggcgggt gaagcgatta
     3361 ccacccatag ttcgcggagg ggccattgac agatactggc ctactgctga tgggcgcctg
     3421 gttgagtatg acatagatga agtggtgtat gatgaagatt caccatatca gaacatcaaa
     3481 atcctacact cgaagcagtt tgggaatatt ctcatcctca gcggggatgt taatttggca
     3541 gagagtgact tggcatatac ccgtgccatc atgggcagtg gtaaagaaga ctacactggc
     3601 aaagatgtac tgattctggg aggcggcgat ggaggcatat tgtgtgaaat tgtcaaactg
     3661 aaaccaaaga tggtcactat ggtagagatt gaccaaatgg taattgatgg atgtaagaaa
     3721 tacatgcgga gaacatgtgg agatgtctta gacaatctta gaggagactg ctatcaggtt
     3781 cttatagaag actgtatccc agtactgaag atgtacgcca aagaagggag agagtttgat
     3841 tatgtgatta atgatttgac agctgtccct atctctacat ctccagaaga agattccaca
     3901 tgggatttcc tcagactgat tctcgacctg tcaatgaagg ttctgaaaca ggatggaaaa
     3961 tacttcacac aggggaactg tgtcaacttg actgaagccc tgtcgctcta tgaagaacag
     4021 ctggggcgtc tgtattgtcc tgtggaattc tcaaaagaga ttgtctgtgt cccttcatac
     4081 ttggaattgt gggtgtttta cactgtttgg aagaaagcta agccttaccc atacgacgta
     4141 ccagattacg cttgaattcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     4201 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     4261 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     4321 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     4381 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     4441 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     4501 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     4561 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     4621 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     4681 agatccaagc tgtgaccggc gcctacgcta ggccaccatg gccaagcctt tgtctcaaga
     4741 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     4801 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     4861 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     4921 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     4981 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     5041 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     5101 tgtgtgggag ggctaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     5161 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5221 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5281 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5341 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5401 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5461 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     5521 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5581 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5641 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5701 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5761 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc
     5821 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5881 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5941 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6001 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     6061 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     6121 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6181 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     6241 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6301 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     6361 aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt
     6421 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6481 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6541 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6601 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6661 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6721 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6781 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6841 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6901 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6961 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7021 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7081 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7141 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7201 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7261 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7321 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7381 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7441 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7501 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7561 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7621 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7681 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7741 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7801 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7861 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7921 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7981 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8041 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8101 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8161 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8221 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8281 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8341 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8401 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8461 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8521 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     8581 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     8641 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     8701 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     8761 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     8821 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     8881 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     8941 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     9001 ttgtaaaacg acggccagtg ccaagctg
//
