LOCUS       V027374                 9622 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027374
VERSION     V027374
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..4713
                     /label="SPTLC2(NM_004863)"
                     /note="SPTLC2(NM_004863)"
                     /gene="SPTLC2"
     CDS             4720..4746
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4787..4998
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5011..5279
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5312..5710
                     /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    5717..6305
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6379..6612
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6684..6805
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6824..6959
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6997..7013)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7021..7037
                     /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(7045..7075)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7090..7111
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7399..7987)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8158..9018)
                     /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(9019..9123)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9597..9613
                     /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 cggccggagc ccggaggctg ctgctgccgc
     3061 cgcacggtgc gggcgaatgg ctgcgtggcg aacggggaag tacggaacgg gtacgtgagg
     3121 agcagcgctg cagccgcagc cgcagccgcc gccggccaga tccatcatgt tacacaaaat
     3181 ggaggactat ataaaagacc gtttaatgaa gcttttgaag aaacaccaat gctggttgct
     3241 gtgctcacgt atgtggggta tggcgtactc accctctttg gatatcttcg agatttcttg
     3301 aggtattgga gaattgaaaa gtgtcaccat gcaacagaaa gagaagaaca aaaggacttt
     3361 gtgtcattgt atcaagattt tgaaaacttt tatacaagga atctgtacat gaggataaga
     3421 gacaactgga atcggccaat ctgtagtgtg cctggagcca gggtggacat catggagaga
     3481 cagtctcatg attataactg gtccttcaag tatacaggga atataataaa gggtgttata
     3541 aacatgggtt cctacaacta tcttggattt gcacggaata ctggatcatg tcaagaagca
     3601 gccgccaaag tccttgagga gtatggagct ggagtgtgca gtactcggca ggaaattgga
     3661 aacctggaca agcatgaaga actagaggag cttgtagcaa ggttcttagg agtagaagct
     3721 gctatggcgt atggcatggg atttgcaacg aattcaatga acattcctgc tcttgttggc
     3781 aaaggttgcc tgattctgag tgatgaactg aatcatgcat cactggttct gggagccaga
     3841 ctgtcaggag caaccattag aatcttcaaa cacaacaata tgcaaagcct agagaagcta
     3901 ttgaaagatg ccattgttta tggtcagcct cggacacgaa ggccctggaa gaaaattctc
     3961 atccttgtgg aaggaatata tagcatggag ggatctattg ttcgtcttcc tgaagtgatt
     4021 gccctcaaga agaaatacaa ggcatacttg tatctggatg aggctcacag cattggcgcc
     4081 ctgggcccca caggccgggg tgtggtggag tactttggcc tggatcccga ggatgtggat
     4141 gttatgatgg gaacgttcac aaagagtttt ggtgcttctg gaggatatat tggaggcaag
     4201 aaggagctga tagactacct gcgaacacat tctcatagtg cagtgtatgc cacgtcattg
     4261 tcacctcctg tagtggagca gatcatcacc tccatgaagt gcatcatggg gcaggatggc
     4321 accagccttg gtaaagagtg tgtacaacag ttagctgaaa acaccaggta tttcaggaga
     4381 cgcctgaaag agatgggctt catcatctat ggaaatgaag actctccagt agtgcctttg
     4441 atgctctaca tgcctgccaa aattggcgcc tttggacggg agatgctgaa gcggaacatc
     4501 ggtgtcgttg tggttggatt tcctgccacc ccaattattg agtccagagc caggttttgc
     4561 ctgtcagcag ctcataccaa agaaatactt gatactgctt taaaggagat agatgaagtt
     4621 ggggacctat tgcagctgaa gtattcccgt catcggttgg tacctctact ggacaggccc
     4681 tttgacgaga cgacgtatga agaaacagaa gacctcgagt acccatacga cgtaccagat
     4741 tacgcttgaa ttcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     4801 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     4861 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     4921 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     4981 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     5041 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     5101 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     5161 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     5221 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     5281 aagctgtgac cggcgcctac gctaggccac catggccaag cctttgtctc aagaagaatc
     5341 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     5401 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     5461 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     5521 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     5581 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
     5641 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     5701 ggagggctaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5761 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5821 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5881 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5941 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6001 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6061 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     6121 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6181 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6241 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6301 cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6361 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt
     6421 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6481 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6541 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6601 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6661 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     6721 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6781 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     6841 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6901 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6961 agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     7021 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7081 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7141 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7201 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7261 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7321 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7381 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7441 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7501 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7561 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7621 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7681 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7741 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7801 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7861 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7921 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7981 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8041 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8101 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8161 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8221 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8281 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8341 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8401 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8461 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8521 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8581 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8641 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8701 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8761 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8821 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8881 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8941 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9001 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9061 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9121 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     9181 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     9241 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     9301 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     9361 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     9421 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     9481 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     9541 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     9601 aacgacggcc agtgccaagc tg
//
