LOCUS       V043964                 9688 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043964
VERSION     V043964
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             2315..4036
                     /label="PKLR(NM_000298)"
                     /note="PKLR(NM_000298)"
                     /gene="PKLR"
     promoter        4081..4292
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4305..4573
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4608..5315
                     /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             5325..5378
                     /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             5379..5777
                     /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    5784..6371
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6445..6678
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6750..6871
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6890..7025
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7063..7079)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7087..7103
                     /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(7111..7141)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7156..7177
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7465..8053)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8224..9084)
                     /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(9085..9189)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9663..9679
                     /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 gaattcgaaa tcggatccgc caccatgtcg atccaggaga acatatcatc
     2341 cctgcagctt cggtcatggg tctctaagtc ccaaagagac ttagcaaagt ccatcctgat
     2401 tggggctcca ggagggccag cggggtatct gcggcgggcc agtgtggccc aactgaccca
     2461 ggagctgggc actgccttct tccagcagca gcagctgcca gctgctatgg cagacacctt
     2521 cctggaacac ctctgcctac tggacattga ctccgagccc gtggctgctc gcagtaccag
     2581 catcattgcc accatcgggc cagcatctcg ctccgtggag cgcctcaagg agatgatcaa
     2641 ggccgggatg aacattgcgc gactcaactt ctcccacggc tcccacgagt accatgctga
     2701 gtccatcgcc aacgtccggg aggcggtgga gagctttgca ggttccccac tcagctaccg
     2761 gcccgtggcc atcgccctgg acaccaaggg accggagatc cgcactggga tcctgcaggg
     2821 gggtccagag tcggaagtgg agctggtgaa gggctcccag gtgctggtga ctgtggaccc
     2881 cgcgttccgg acgcggggga acgcgaacac cgtgtgggtg gactacccca atattgtccg
     2941 ggtcgtgccg gtggggggcc gcatctacat tgacgacggg ctcatctccc tagtggtcca
     3001 gaaaatcggc ccagagggac tggtgaccca agtggagaac ggcggcgtcc tgggcagccg
     3061 gaagggcgtg aacttgccag gggcccaggt ggacttgccc gggctgtccg agcaggacgt
     3121 ccgagacctg cgcttcgggg tggagcatgg ggtggacatc gtctttgcct cctttgtgcg
     3181 gaaagccagc gacgtggctg ccgtcagggc tgctctgggt ccggaaggac acggcatcaa
     3241 gatcatcagc aaaattgaga accacgaagg cgtgaagagg tttgatgaaa tcctggaggt
     3301 gagcgacggc atcatggtgg cacgggggga cctaggcatc gagatcccag cagagaaggt
     3361 tttcctggct cagaagatga tgattgggcg ctgcaacttg gcgggcaagc ctgttgtctg
     3421 tgccacacag atgctggaga gcatgattac caagccccgg ccaacgaggg cagagacaag
     3481 cgatgtcgcc aatgctgtgc tggatggggc tgactgcatc atgctgtcag gggagactgc
     3541 caagggcaac ttccctgtgg aagcggtgaa gatgcagcat gcgattgccc gggaggcaga
     3601 ggccgcagtg taccaccggc agctgtttga ggagctacgt cgggcagcgc cactaagccg
     3661 tgatcccact gaggtcaccg ccattggtgc tgtggaggct gccttcaagt gctgtgctgc
     3721 tgccatcatt gtgctgacca caactggccg ctcagcccag cttctgtctc ggtaccgacc
     3781 tcgggcagca gtcattgctg tcacccgctc tgcccaggct gcccgccagg tccacttatg
     3841 ccgaggagtc ttccccttgc tttaccgtga acctccagaa gccatctggg cagatgatgt
     3901 agatcgccgg gtgcaatttg gcattgaaag tggaaagctc cgtggcttcc tccgtgttgg
     3961 agacctggtg attgtggtga caggctggcg acctggctcc ggctacacca acatcatgcg
     4021 ggtgctaagc atatcctgag cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt
     4081 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     4141 cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     4201 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     4261 tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc
     4321 cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc
     4381 cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca
     4441 ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct
     4501 ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc
     4561 gccgttacag atccaagctg tgaccggcgc ctacgctaga cgccaccatg gtgagcaagg
     4621 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     4681 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     4741 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     4801 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     4861 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     4921 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     4981 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     5041 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     5101 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     5161 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     5221 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     5281 gccactccac cggcggcatg gacgagctgt acaagggatc tcgcgagggc agaggaagtc
     5341 ttctaacatg cggtgacgtg gaggagaatc ccggccctat ggccaagcct ttgtctcaag
     5401 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     5461 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     5521 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     5581 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     5641 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     5701 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     5761 atgtgtggga gggctaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     5821 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     5881 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     5941 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     6001 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     6061 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6121 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     6181 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6241 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6301 ctctgcggcc tcttccgcgt ctccgccttc gccctcagac gagtcggatc tccctttggc
     6361 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     6421 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agataagatc
     6481 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6541 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6601 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6661 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     6721 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     6781 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6841 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     6901 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6961 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     7021 aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt
     7081 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     7141 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7201 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7261 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7321 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7381 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7441 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7501 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7561 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7621 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7681 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7741 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7801 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7861 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7921 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7981 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     8041 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8101 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8161 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8221 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8281 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8341 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8401 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8461 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8521 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8581 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8641 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8701 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8761 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8821 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8881 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8941 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     9001 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9061 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     9121 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9181 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     9241 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     9301 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     9361 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     9421 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     9481 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     9541 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     9601 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     9661 ttgtaaaacg acggccagtg ccaagctg
//
