LOCUS       V040814                 9669 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040814
VERSION     V040814
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3946
                     /label="LGI3(NM_139278)"
                     /note="LGI3(NM_139278)"
                     /gene="LGI3"
     CDS             3953..4018
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4062..4273
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4286..4554
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4589..5296
                     /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             5306..5359
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             5360..5758
                     /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    5765..6352
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6426..6659
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6731..6852
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6871..7006
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7044..7060)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7068..7084
                     /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(7092..7122)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7137..7158
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7446..8034)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8205..9065)
                     /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(9066..9170)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9644..9660
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatggcggg gctgcgggcc agggggggcc cggggccggg
     2341 gctgctggcg ctctccgcgc tcggcttctg cctcatgctg caagtcagcg ctaagaggcc
     2401 ccccaagacg cccccctgcc cgcccagctg ctcttgcacc agggacaccg ccttctgcgt
     2461 ggactcaaag gcggtgccca ggaacctgcc ctcggaggtc atctccctga ccctggtgaa
     2521 tgccgccttc tcagagatcc aggatggagc gttctcccac ctcccgctgc tgcagttctt
     2581 gttactcaac tccaacaagt ttacactgat tggagacaac gccttcacag gactgtcgca
     2641 cctgcagtat ctcttcattg agaacaatga catctgggca ctatccaagt tcaccttccg
     2701 aggactcaag tccttgactc acctctcgct ggccaacaat aacctgcaga cactgcccag
     2761 agacatcttc cggcccctgg acatcctgaa tgacttggac ctgcggggca actcactcaa
     2821 ctgtgactgc aaggtgaagt ggttggtgga gtggctggca cacaccaaca ccacggtggc
     2881 acccatctac tgcgccagcc cgccccgctt ccaggagcac aaggtgcagg acctgccgct
     2941 gcgggagttc gattgcatca ccacagattt tgtgttgtac cagaccctgg ccttcccagc
     3001 agtgtcggct gagcccttcc tctactccag tgacctctat ttggctctgg cccagccagg
     3061 agtcagtgcc tgcaccatcc tgaagtggga ctatgttgag cggcagcttc gagactatga
     3121 tagaatccca gccccctctg cagtgcactg caagccgatg gtggtggaca gccagctgta
     3181 cgtggtcgtg gcccagctgt ttggcggctc ttacatttac cactgggatc ccaacaccac
     3241 gcgcttcacc aggctgcaag acattgaccc gcagcgcgtg cgcaagccta acgacctaga
     3301 agccttccgc atcgacggtg actggtactt tgccgtggct gacagctcca aggcaggcgc
     3361 caccagcctc taccgctggc accagaatgg cttctactcc caccaggcac tgcacccctg
     3421 gcaccgtgac accgacctgg agtttgtgga cggcgagggc aagccacggc tgattgtgtc
     3481 cagcagctcc caggcacccg tcatctatca gtggagtcgc acccagaagc agtttgtggc
     3541 ccagggtgag gtgacccagg tgcctgatgc ccaagctgtg aaacactttc gtgccggccg
     3601 cgacagctac ctgtgcctca gccgctacat tggcgactcc aagatcctgc gctgggaggg
     3661 tacccgcttc tcggaggtgc aggccctgcc ctcccggggc tcgctggccc tgcagccctt
     3721 ccttgtgggt ggccgccgct acctggcact gggcagtgat ttctccttca cccagatcta
     3781 ccagtgggat gagggacgac agaagtttgt acggttccag gagctggctg tgcaggctcc
     3841 tcgggccttc tgctacatgc ctgctgggga cgcccagcta ctcctggccc ccagcttcaa
     3901 gggacagacg ctggtgtata gacacattgt ggtggatctc agtgccggat ccgactacaa
     3961 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     4021 gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc
     4081 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg
     4141 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     4201 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     4261 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc
     4321 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
     4381 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
     4441 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
     4501 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
     4561 gtgaccggcg cctacgctag acgccaccat ggtgagcaag ggcgaggagg ataacatggc
     4621 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     4681 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     4741 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     4801 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     4861 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     4921 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     4981 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     5041 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     5101 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     5161 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     5221 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     5281 ggacgagctg tacaagggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt
     5341 ggaggagaat cccggcccta tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     5401 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     5461 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     5521 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     5581 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     5641 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     5701 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt
     5761 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5821 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5881 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5941 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     6001 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     6061 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     6121 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     6181 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6241 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     6301 tctccgcctt cgccctcaga cgagtcggat ctccctttgg ccgcctcccc gcctggtacc
     6361 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     6421 gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg
     6481 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     6541 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     6601 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     6661 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     6721 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6781 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6841 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc
     6901 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     6961 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga
     7021 gacggcccaa attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     7081 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     7141 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     7201 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     7261 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     7321 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     7381 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     7441 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7501 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7561 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7621 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7681 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7741 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7801 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7861 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     7921 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7981 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     8041 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     8101 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     8161 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     8221 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     8281 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     8341 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     8401 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     8461 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     8521 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     8581 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8641 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8701 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8761 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8821 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8881 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8941 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     9001 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     9061 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     9121 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     9181 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     9241 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     9301 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     9361 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     9421 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     9481 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     9541 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     9601 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     9661 gccaagctg
//
