LOCUS       V043030                10018 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043030
VERSION     V043030
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..4671
                     /label="CDH6(NM_004932)"
                     /note="CDH6(NM_004932)"
                     /gene="CDH6"
     CDS             4678..4743
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4787..4998
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5011..5279
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5312..6106
                     /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    6113..6701
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6775..7008
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7080..7201
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7220..7355
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7393..7409)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7417..7433
                     /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(7441..7471)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7486..7507
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7795..8383)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8554..9414)
                     /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(9415..9519)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9993..10009
                     /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 catgagaact taccgctact tcttgctgct cttttgggtg
     2341 ggccagccct acccaactct ctcaactcca ctatcaaaga ggactagtgg tttcccagca
     2401 aagaaaaggg ccctggagct ctctggaaac agcaaaaatg agctgaaccg ttcaaaaagg
     2461 agctggatgt ggaatcagtt ctttctcctg gaggaataca caggatccga ttatcagtat
     2521 gtgggcaagt tacattcaga ccaggataga ggagatggat cacttaaata tatcctttca
     2581 ggagatggag caggagatct cttcattatt aatgaaaaca caggcgacat acaggccacc
     2641 aagaggctgg acagggaaga aaaacccgtt tacatccttc gagctcaagc tataaacaga
     2701 aggacaggga gacccgtgga gcccgagtct gaattcatca tcaagatcca tgacatcaat
     2761 gacaatgaac caatattcac caaggaggtt tacacagcca ctgtccctga aatgtctgat
     2821 gtcggtacat ttgttgtcca agtcactgcg acggatgcag atgatccaac atatgggaac
     2881 agtgctaaag ttgtctacag tattctacag ggacagccct atttttcagt tgaatcagaa
     2941 acaggtatta tcaagacagc tttgctcaac atggatcgag aaaacaggga gcagtaccaa
     3001 gtggtgattc aagccaagga tatgggcggc cagatgggag gattatctgg gaccaccacc
     3061 gtgaacatca cactgactga tgtcaacgac aaccctcccc gattccccca gagtacatac
     3121 cagtttaaaa ctcctgaatc ttctccaccg gggacaccaa ttggcagaat caaagccagc
     3181 gacgctgatg tgggagaaaa tgctgaaatt gagtacagca tcacagacgg tgaggggctg
     3241 gatatgtttg atgtcatcac cgaccaggaa acccaggaag ggattataac tgtcaaaaag
     3301 ctcttggact ttgaaaagaa gaaagtgtat acccttaaag tggaagcctc caatccttat
     3361 gttgagccac gatttctcta cttggggcct ttcaaagatt cagccacggt tagaattgtg
     3421 gtggaggatg tagatgagcc acctgtcttc agcaaactgg cctacatctt acaaataaga
     3481 gaagatgctc agataaacac cacaataggc tccgtcacag cccaagatcc agatgctgcc
     3541 aggaatcctg tcaagtactc tgtagatcga cacacagata tggacagaat attcaacatt
     3601 gattctggaa atggttcgat ttttacatcg aaacttcttg accgagaaac actgctatgg
     3661 cacaacatta cagtgatagc aacagagatc aataatccaa agcaaagtag tcgagtacct
     3721 ctatatatta aagttctaga tgtcaatgac aacgccccag aatttgctga gttctatgaa
     3781 acttttgtct gtgaaaaagc aaaggcagat cagttgattc agaccctgca tgctgttgac
     3841 aaggatgacc cttatagtgg acaccaattt tcgttttcct tggcccctga agcagccagt
     3901 ggctcaaact ttaccattca agacaacaaa gacaacacgg cgggaatctt aactcggaaa
     3961 aatggctata atagacacga gatgagcacc tatctcttgc ctgtggtcat ttcagacaac
     4021 gactacccag ttcaaagcag cactgggaca gtgactgtcc gggtctgtgc atgtgaccac
     4081 cacgggaaca tgcaatcctg ccatgcggag gcgctcatcc accccacggg actgagcacg
     4141 ggggctctgg ttgccatcct tctgtgcatc gtgatcctac tagtgacagt ggtgctgttt
     4201 gcagctctga ggcggcagcg aaaaaaagag cctttgatca tttccaaaga ggacatcaga
     4261 gataacattg tcagttacaa cgacgaaggt ggtggagagg aggacaccca ggcttttgat
     4321 atcggcaccc tgaggaatcc tgaagccata gaggacaaca aattacgaag ggacattgtg
     4381 cccgaagccc ttttcctacc ccgacggact ccaacagctc gcgacaacac cgatgtcaga
     4441 gatttcatta accaaaggtt aaaggaaaat gacacggacc ccactgcccc gccatacgac
     4501 tccttggcca cttacgccta tgaaggcact ggctccgtgg cggattccct gagctcgctg
     4561 gagtcagtga ccacggatgc agatcaagac tatgattacc ttagtgactg gggacctcga
     4621 ttcaaaaagc ttgcagatat gtatggagga gtggacagtg acaaagactc cggatccgac
     4681 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     4741 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     4801 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     4861 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     4921 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     4981 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     5041 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     5101 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     5161 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     5221 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     5281 aagctgtgac cggcgcctac gctaggccac catggctgaa caagatggat tgcacgcagg
     5341 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5401 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5461 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5521 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5581 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5641 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5701 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5761 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5821 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5881 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5941 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6001 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6061 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtcg acaatcaacc
     6121 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     6181 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     6241 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     6301 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     6361 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     6421 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     6481 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     6541 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6601 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6661 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa
     6721 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6781 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt
     6841 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6901 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6961 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     7021 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     7081 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     7141 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7201 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     7261 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     7321 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa
     7381 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7441 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7501 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7561 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7621 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7681 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7741 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7801 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7861 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7921 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7981 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8041 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8101 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8161 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8221 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8281 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8341 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8401 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8461 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8521 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8581 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8641 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8701 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8761 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8821 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8881 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8941 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9001 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9061 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9121 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9181 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9241 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9301 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9361 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9421 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9481 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9541 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     9601 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     9661 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     9721 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     9781 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     9841 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     9901 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     9961 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
