LOCUS       V041453                 9008 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041453
VERSION     V041453
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             2302..3699
                     /label="EED(NM_001308007)"
                     /note="EED(NM_001308007)"
                     /gene="EED"
     CDS             3706..3732
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        3777..3988
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4001..4269
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4302..5096
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5103..5691
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5765..5998
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6070..6191
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6210..6345
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6383..6399)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6407..6423
                     /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(6431..6461)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6476..6497
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6785..7373)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7544..8404)
                     /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(8405..8509)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8983..8999
                     /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 agaagattct
     2281 agagctagcg aattcgccac catgtccgag agggaagtgt cgactgcgcc ggcgggaaca
     2341 gacatgcctg cggccaagaa gcagaagctg agcagtgacg agaacagcaa tccagacctc
     2401 tctggagacg agaatgatga cgctgtcagt atagaaagtg gtacaaacac tgaacgccct
     2461 gatacaccta caaacacgcc aaatgcacct ggaaggaaaa gttggggaaa gggaaaatgg
     2521 aagtcaaaga aatgcaaata ttctttcaaa tgtgtaaata gtctcaagga agatcataac
     2581 caaccattgt ttggagttca gtttaactgg cacagtaaag aaggagatcc attagtgttt
     2641 gcaactgtag gaagcaacag agttaccttg tatgaatgtc attcacaagg agaaatccgg
     2701 ttgttgcaat cttacgtgga tgctgatgct gatgaaaact tttacacttg tgcatggacc
     2761 tatgatagca atacgagcca tcctctgctg gctgtagctg gatctagagg cataattagg
     2821 ataataaatc ctataacaat gcagtgtata aagcactatg ttggccatgg aaatgctatc
     2881 aatgagctga aattccatcc aagagatcca aatcttctcc tgtcagtaag taaagatcat
     2941 gctttacgat tatggaatat ccagacggac actctggtgg caatatttgg aggcgtagaa
     3001 gggcacagag atgaagttct aagtgctgat tatgatcttt tgggtgaaaa aataatgtcc
     3061 tgtggtatgg atcattctct taaactttgg aggatcaatt caaagagaat gatgaatgca
     3121 attaaggaat cttatgatta taatccaaat aaaactaaca ggccatttat ttctcagaaa
     3181 atccattttc ctgatttttc taccagagac atacatagga attatgttga ttgtgtgcga
     3241 tggttaggcg atttgatact ttctaagagt ggccgtgcca ttttacattc ccaccagcaa
     3301 tgtatgagag atccagtgtc tccgaatctt cgccagcatt tgtcttgtga aaatgccatt
     3361 gtgtgctgga aacctggcaa gatggaagat gatatagata aaattaaacc cagtgaatct
     3421 aatgtgacta ttcttgggcg atttgattac agccagtgtg acatttggta catgaggttt
     3481 tctatggatt tctggcaaaa gatgcttgca ttgggcaatc aagttggcaa actttatgtt
     3541 tgggatttag aagtagaaga tcctcataaa gccaaatgta caacactgac tcatcataaa
     3601 tgtggtgctg ctattcgaca aaccagtttt agcagggata gcagcattct tatagctgtt
     3661 tgtgatgatg ccagtatttg gcgctgggat cgacttcgag gatcctaccc atacgatgtt
     3721 ccagattacg cttaggcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     3781 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     3841 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     3901 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     3961 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     4021 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     4081 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     4141 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     4201 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     4261 ttacagatcc aagctgtgac cggcgcctac gctaggccac catggctgaa caagatggat
     4321 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4381 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4441 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4501 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4561 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4621 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4681 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4741 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4801 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4861 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4921 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4981 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5041 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtcg
     5101 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5161 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5221 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5281 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5341 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5401 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5461 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     5521 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5581 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5641 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct
     5701 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     5761 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg
     5821 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     5881 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     5941 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6001 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     6061 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6121 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6181 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc
     6241 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6301 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag
     6361 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6421 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6481 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6541 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6601 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6661 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6721 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6781 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6841 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6901 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6961 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7021 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7081 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7141 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7201 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7261 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7321 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7381 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7441 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7501 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7561 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7621 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7681 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7741 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7801 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7861 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7921 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7981 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8041 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8101 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8161 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8221 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8281 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8341 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8401 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8461 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8521 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     8581 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     8641 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     8701 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     8761 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     8821 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     8881 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     8941 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     9001 ccaagctg
//
