LOCUS       V040415                 8920 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040415
VERSION     V040415
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..3549
                     /label="MKRN2(NM_014160)"
                     /note="MKRN2(NM_014160)"
                     /gene="MKRN2"
     CDS             3556..3645
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3689..3900
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3913..4181
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4214..5008
                     /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    5015..5603
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5677..5910
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5982..6103
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6122..6257
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6295..6311)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6319..6335
                     /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(6343..6373)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6388..6409
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6697..7285)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7456..8316)
                     /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(8317..8421)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8895..8911
                     /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 catgagcacc aagcagatca cttgcaggta ttttatgcat
     2341 ggtgtgtgtc gggaaggaag tcagtgccta ttctcacatg acttggcaaa cagcaaaccg
     2401 tccaccatct gcaagtacta ccagaagggc tactgtgcct atggaactcg gtgcagatat
     2461 gaccacacga ggccctctgc tgcagctgga ggtgctgtgg gcaccatggc ccacagtgtg
     2521 ccctccccag ctttccacag tcctcaccct ccttccgagg tcactgcatc cattgtgaaa
     2581 actaactcac atgaacccgg aaagcgtgaa aagagaacat tggttcttag agaccgaaat
     2641 ctctctggca tggctgaaag gaagacccag ccgagcatgg tgagtaatcc aggcagctgc
     2701 agcgaccccc agcccagccc cgagatgaag ccgcattcct acctggatgc catcaggagt
     2761 ggccttgatg acgtggaggc cagcagctcc tacagcaacg agcagcagct gtgcccctac
     2821 gcagctgctg gggagtgccg gtttggggat gcctgtgtct acctgcacgg ggaggtgtgt
     2881 gaaatctgta ggctgcaagt cttgcaccca ttcgacccag agcagaggaa ggctcatgaa
     2941 aagatctgca tgttgacgtt cgaacacgag atggaaaagg cctttgcctt ccaggcaagc
     3001 caggacaaag tgtgcagtat ctgcatggaa gtgatcctgg agaaggcctc tgcttctgag
     3061 aggagatttg ggattctctc caattgcaat cacacgtact gtttgtcctg catccggcag
     3121 tggcggtgtg ccaaacagtt tgaaaaccca atcattaagt cttgtccaga atgccgtgtg
     3181 atatcagagt ttgtaattcc aagtgtgtat tgggtggaag atcagaataa aaagaacgag
     3241 ttgattgaag ctttcaaaca ggggatgggg aaaaaagcct gtaaatactt tgagcaaggc
     3301 aaggggacct gcccatttgg aagcaaatgt ctttatcgcc atgcttaccc cgatgggcgg
     3361 ctagcagagc ctgagaaacc tcggaaacag ctcagttctc aaggcactgt gaggttcttt
     3421 aattcagtgc ggctctggga tttcatcgag aaccgagaaa gccggcatgt ccccaacaat
     3481 gaagatgtcg acatgacaga gctcggggac ctcttcatgc acctttctgg agtggaatca
     3541 tcagaacccg gatccgaaca aaaactaata tcagaggaag atttggaaca gaaactaatc
     3601 agtgaagaag atttagagca aaagttaatt tccgaggagg acttatagcg gccgcgaagg
     3661 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     3721 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac
     3781 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     3841 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     3901 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc
     3961 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg
     4021 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct
     4081 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc
     4141 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct
     4201 acgctaggcc accatggctg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4261 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4321 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4381 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4441 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4501 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4561 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4621 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4681 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4741 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4801 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4861 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4921 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4981 ctatcgcctt cttgacgagt tcttctgagt cgacaatcaa cctctggatt acaaaatttg
     5041 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5101 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5161 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5221 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5281 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5341 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5401 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     5461 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5521 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5581 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     5641 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     5701 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     5761 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     5821 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     5881 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     5941 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6001 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6061 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     6121 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6181 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6241 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     6301 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6361 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6421 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6481 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6541 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6601 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6661 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6721 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6781 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6841 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6901 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6961 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7021 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7081 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7141 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7201 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7261 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7321 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7381 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7441 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7501 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7561 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7621 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7681 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7741 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7801 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7861 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7921 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7981 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8041 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8101 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8161 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8221 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8281 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8341 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8401 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     8461 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     8521 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     8581 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     8641 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     8701 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     8761 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     8821 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     8881 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
