LOCUS       V043726                 9556 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043726
VERSION     V043726
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..3904
                     /label="FDXR(NM_001258012)"
                     /note="FDXR(NM_001258012)"
                     /gene="FDXR"
     promoter        3949..4160
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4173..4441
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4476..5183
                     /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             5193..5246
                     /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             5247..5645
                     /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    5652..6239
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6313..6546
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6618..6739
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6758..6893
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6931..6947)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6955..6971
                     /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(6979..7009)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7024..7045
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7333..7921)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8092..8952)
                     /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(8953..9057)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9531..9547
                     /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 ccatggcttc gcgctgctgg cgctggtggg gctggtcggc
     2341 gtggcctcgg acccggctgc ctcccgccgg gagcaccccg agcttctgcc accatttctc
     2401 cacacaggag aagacccccc agatctgtgt ggtgggcagt ggcccagctg gcttctacac
     2461 ggcccaacac ctgctaaaga gggtggaagc cttgtgttct cagcccaggg tcctgaactc
     2521 tcctgctctg tctggggaag gggaggacct gggggcgtcc cagcctctct ctctcggccc
     2581 caccagctgc caccctgttc cccagcagca cccccaggcc cacgtggaca tctacgagaa
     2641 acagcctgtg ccctttggcc tggtgcgctt tggtgtggcg cctgatcacc ccgaggtgaa
     2701 gaatgtcatc aacacattta cccagacggc ccattctggc cgctgtgcct tctggggcaa
     2761 cgtggaggtg ggcagggacg tgacggtgcc ggagctgcag gaggcctacc acgctgtggt
     2821 gctgagctac ggggcagagg accatcgggc cctggaaatt cctggtgagg agctgccagg
     2881 tgtgtgctcc gcccgggcct tcgtgggctg gtacaacggg cttcctgaga accaggagct
     2941 ggagccagac ctgagctgtg acacagccgt gattctgggg caggggaacg tggctctgga
     3001 cgtggcccgc atcctactga ccccacctga gcacctggag agaacggaca tcacgaaggc
     3061 agccctgggt gtactgaggc agagtcgagt gaagacagtg tggctagtgg gccggcgtgg
     3121 acccctgcaa gtggccttca ccattaagga gcttcgggag atgattcagt taccgggagc
     3181 ccggcccatt ttggatcctg tggatttctt gggtctccag gacaagatca aggaggtccc
     3241 ccgcccgagg aagcggctga cggaactgct gcttcgaacg gccacagaga agccagggcc
     3301 ggcggaagct gcccgccagg catcggcctc ccgtgcctgg ggcctccgct ttttccgaag
     3361 cccccagcag gtgctgccct caccagatgg gcggcgggca gcaggtgtcc gcctagcagt
     3421 cactagactg gagggtgtcg atgaggccac ccgtgcagtg cccacgggag acatggaaga
     3481 cctcccttgt gggctggtgc tcagcagcat tgggtataag agccgccctg tcgacccaag
     3541 cgtgcccttt gactccaagc ttggggtcat ccccaatgtg gagggccggg ttatggatgt
     3601 gccaggcctc tactgcagcg gctgggtgaa gagaggacct acaggtgtca tagccacaac
     3661 catgactgac agcttcctca ccggccagat gctgctgcag gacctgaagg ctgggttgct
     3721 cccctctggc cccaggcctg gctacgcagc catccaggcc ctgctcagca gccgaggggt
     3781 ccggccagtc tctttctcag actgggagaa gctggatgcc gaggaggtgg cccggggcca
     3841 gggcacgggg aagcccaggg agaagctggt ggatcctcag gagatgctgc gcctcctggg
     3901 ccactgagcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca
     3961 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga
     4021 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     4081 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     4141 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc
     4201 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct
     4261 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg
     4321 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc
     4381 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat
     4441 ccaagctgtg accggcgcct acgctagacg ccaccatggt gagcaagggc gaggaggata
     4501 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     4561 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     4621 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     4681 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     4741 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     4801 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     4861 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     4921 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     4981 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     5041 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     5101 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     5161 gcggcatgga cgagctgtac aagggatctc gcgagggcag aggaagtctt ctaacatgcg
     5221 gtgacgtgga ggagaatccc ggccctatgg ccaagccttt gtctcaagaa gaatccaccc
     5281 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     5341 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     5401 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     5461 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     5521 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     5581 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     5641 gctaagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5701 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5761 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5821 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5881 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5941 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6001 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     6061 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6121 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6181 ttccgcgtct ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc
     6241 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     6301 aaaagggggg actggaaggg ctaattcact cccaacgaag ataagatctg ctttttgctt
     6361 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6421 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6481 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6541 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     6601 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6661 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6721 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag
     6781 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6841 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     6901 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6961 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7021 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7081 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7141 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7201 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7261 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7741 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7921 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7981 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8041 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8101 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8161 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8221 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8281 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8341 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8401 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8461 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8521 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8581 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8641 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8701 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8761 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8821 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8881 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8941 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9001 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9061 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     9121 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     9181 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     9241 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     9301 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     9361 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     9421 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     9481 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     9541 ggccagtgcc aagctg
//
