LOCUS       V042473                10138 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042473
VERSION     V042473
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..4839
                     /label="EXO1(NM_006027)"
                     /note="EXO1(NM_006027)"
                     /gene="EXO1"
     CDS             4840..4863
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4907..5118
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5131..5399
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5432..6226
                     /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    6233..6821
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6895..7128
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7200..7321
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7340..7475
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7513..7529)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7537..7553
                     /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(7561..7591)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7606..7627
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7915..8503)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8674..9534)
                     /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(9535..9639)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10113..10129
                     /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 catggggata cagggattgc tacaatttat caaagaagct
     2341 tcagaaccca tccatgtgag gaagtataaa gggcaggtag tagctgtgga tacatattgc
     2401 tggcttcaca aaggagctat tgcttgtgct gaaaaactag ccaaaggtga acctactgat
     2461 aggtatgtag gattttgtat gaaatttgta aatatgttac tatctcatgg gatcaagcct
     2521 attctcgtat ttgatggatg tactttacct tctaaaaagg aagtagagag atctagaaga
     2581 gaaagacgac aagccaatct tcttaaggga aagcaacttc ttcgtgaggg gaaagtctcg
     2641 gaagctcgag agtgtttcac ccggtctatc aatatcacac atgccatggc ccacaaagta
     2701 attaaagctg cccggtctca gggggtagat tgcctcgtgg ctccctatga agctgatgcg
     2761 cagttggcct atcttaacaa agcgggaatt gtgcaagcca taattacaga ggactcggat
     2821 ctcctagctt ttggctgtaa aaaggtaatt ttaaagatgg accagtttgg aaatggactt
     2881 gaaattgatc aagctcggct aggaatgtgc agacagcttg gggatgtatt cacggaagag
     2941 aagtttcgtt acatgtgtat tctttcaggt tgtgactacc tgtcatcact gcgtgggatt
     3001 ggattagcaa aggcatgcaa agtcctaaga ctagccaata atccagatat agtaaaggtt
     3061 atcaagaaaa ttggacatta tctcaagatg aatatcacgg taccagagga ttacatcaac
     3121 gggtttattc gggccaacaa taccttcctc tatcagctag tttttgatcc catcaaaagg
     3181 aaacttattc ctctgaacgc ctatgaagat gatgttgatc ctgaaacact aagctacgct
     3241 gggcaatatg ttgatgattc catagctctt caaatagcac ttggaaataa agatataaat
     3301 acttttgaac agatcgatga ctacaatcca gacactgcta tgcctgccca ttcaagaagt
     3361 catagttggg atgacaaaac atgtcaaaag tcagctaatg ttagcagcat ttggcatagg
     3421 aattactctc ccagaccaga gtcgggtact gtttcagatg ccccacaatt gaaggaaaat
     3481 ccaagtactg tgggagtgga acgagtgatt agtactaaag ggttaaatct cccaaggaaa
     3541 tcatccattg tgaaaagacc aagaagtgca gagctgtcag aagatgacct gttgagtcag
     3601 tattctcttt catttacgaa gaagaccaag aaaaatagct ctgaaggcaa taaatcattg
     3661 agcttttctg aagtgtttgt gcctgacctg gtaaatggac ctactaacaa aaagagtgta
     3721 agcactccac ctaggacgag aaataaattt gcaacatttt tacaaaggaa aaatgaagaa
     3781 agtggtgcag ttgtggttcc agggaccaga agcaggtttt tttgcagttc agattctact
     3841 gactgtgtat caaacaaagt gagcatccag cctctggatg aaactgctgt cacagataaa
     3901 gagaacaatc tgcatgaatc agagtatgga gaccaagaag gcaagagact ggttgacaca
     3961 gatgtagcac gtaattcaag tgatgacatt ccgaataatc atattccagg tgatcatatt
     4021 ccagacaagg caacagtgtt tacagatgaa gagtcctact cttttgagag cagcaaattt
     4081 acaaggacca tttcaccacc cactttggga acactaagaa gttgttttag ttggtctgga
     4141 ggtcttggag atttttcaag aacgccgagc ccctctccaa gcacagcatt gcagcagttc
     4201 cgaagaaaga gcgattcccc cacctctttg cctgagaata atatgtctga tgtgtcgcag
     4261 ttaaagagcg aggagtccag tgacgatgag tctcatccct tacgagaaga ggcatgttct
     4321 tcacagtccc aggaaagtgg agaattctca ctgcagagtt caaatgcatc aaagctttct
     4381 cagtgctcta gtaaggactc tgattcagag gaatctgatt gcaatattaa gttacttgac
     4441 agtcaaagtg accagacctc caagctacgt ttatctcatt tctcaaaaaa agacacacct
     4501 ctaaggaaca aggttcctgg gctatataag tccagttctg cagactctct ttctacaacc
     4561 aagatcaaac ctctaggacc tgccagagcc agtgggctga gcaagaagcc ggcaagcatc
     4621 cagaagagaa agcatcataa tgccgagaac aagccggggt tacagatcaa actcaatgag
     4681 ctctggaaaa actttggatt taaaaaagat tctgaaaagc ttcctccttg taagaaaccc
     4741 ctgtccccag tcagagataa catccaacta actccagaag cggaagagga tatatttaac
     4801 aaacctgaat gtggccgtgt tcaaagagca atattccagg attacaagga cgacgatgac
     4861 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     4921 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     4981 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     5041 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     5101 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     5161 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     5221 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     5281 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     5341 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     5401 aagctgtgac cggcgcctac gctaggccac catggctgaa caagatggat tgcacgcagg
     5461 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5521 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5581 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5641 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5701 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5761 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5821 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5881 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5941 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6001 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     6061 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6121 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6181 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtcg acaatcaacc
     6241 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     6301 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     6361 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     6421 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     6481 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     6541 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     6601 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     6661 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6721 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6781 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa
     6841 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6901 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt
     6961 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     7021 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     7081 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     7141 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     7201 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     7261 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7321 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     7381 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     7441 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa
     7501 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7561 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7621 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7681 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7741 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7801 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7861 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7921 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7981 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8041 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8101 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8161 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8221 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8281 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8341 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8401 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8461 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8521 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8581 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8641 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8701 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8761 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8821 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8881 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8941 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9001 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9061 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9121 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9181 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9241 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9301 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9361 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9421 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9481 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9541 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9601 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9661 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     9721 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     9781 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     9841 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     9901 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     9961 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
    10021 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
    10081 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
