LOCUS       V044568                 9265 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044568
VERSION     V044568
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..3918
                     /label="FBXL6(NM_012162)"
                     /note="FBXL6(NM_012162)"
                     /gene="FBXL6"
     CDS             3925..3990
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4034..4245
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4258..4526
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4559..5353
                     /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    5360..5948
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6022..6255
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6327..6448
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6467..6602
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6640..6656)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6664..6680
                     /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(6688..6718)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6733..6754
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7042..7630)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7801..8661)
                     /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(8662..8766)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9240..9256
                     /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 catggctgcc ccagcctccc ggcaggtccg acgcagagcc
     2341 cgggcagcgc cgcggccccg ctcggccgag gactggtggt gggaccggct ggcgccgagg
     2401 ggctcggggt accacctgct gcagtccgac agcatgctgc tggtgctgtc cgaacccggc
     2461 cccgcccggc cccgcgcaca gcggcgcgct tcccgccgca ctccccggca gccgccccgg
     2521 ggccccagcg ccgcggccaa gcccaaggcc gggctcaggt ccgaggcggc ggccgcgccc
     2581 gcacccgcac cggcacccac gcccacgccc gaggaagggc ccgacgcggg ctggggagac
     2641 cgcattccct tggaaatcct ggtgcagatt ttcgggttgt tggtggcggc ggacggcccc
     2701 atgcccttcc tgggcagggc tgcgcgcgtg tgccgccgct ggcaggaggc cgcttcccaa
     2761 cccgcgctct ggcacaccgt gaccctgtcg tccccgctgg tcggccggcc tgccaagggc
     2821 ggggtcaagg cggagaagaa gctccttgct tccctggagt ggcttatgcc caatcggttt
     2881 tcacagctcc agaggctgac cctcatccac tggaagtctc aggtacaccc cgtgttgaag
     2941 ctggtaggtg agtgctgtcc tcggctcact ttcctcaagc tctccggctg ccacggtgtg
     3001 actgctgacg ctctggtcat gctagccaaa gcctgctgcc agctccatag cctggaccta
     3061 cagcactcca tggtggagtc cacagctgtg gtgagcttct tggaggaggc agggtcccga
     3121 atgcgcaagt tgtggctgac ctacagctcc cagacgacag ccatcctggg cgcactgctg
     3181 ggcagctgct gcccccagct ccaggtcctg gaggtgagca ccggcatcaa ccgtaatagc
     3241 attccccttc agctgcctgt cgaggctctg cagaaaggct gccctcagct ccaggtgctg
     3301 cggctgttga acctgatgtg gctgcccaag cctccgggac gaggggtggc tcccggacca
     3361 ggcttcccta gcctagagga gctctgcctg gcgagctcaa cctgcaactt tgtgagcaac
     3421 gaggtcctgg gccgcctact ccacggctct cccaacctgc gcttactgga tcttcgtggc
     3481 tgtgcgcgca tcacgccggc tggccttcag gatctgccat gtcgggagct ggagcagctt
     3541 catctgggcc tgtatggcac gtcagaccgg ctgactctag ccaaggaggg cagccccttt
     3601 ttgacccaga agtggtgcca tacactgcga gaactggact tgagtggcca ggggttcagt
     3661 gagaaggacc tggagcaggc cctggctgcc ttcttaagca cccctggggg ctcacaccca
     3721 gccctgtgct ctcttaacct caggggcacc cgggtcacac caagcactgt cagctctgtg
     3781 atcagcggct gcccgggcct gctctacctc aacctggagt cctgccgctg ccttccccgg
     3841 ggtctgaagc gggcctaccg gggcctggag gaagtccagt ggtgtctgga gcagctgctc
     3901 accagcccct cacccagcgg atccgactac aaagaccatg acggtgatta taaagatcat
     3961 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc
     4021 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     4081 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     4141 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     4201 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     4261 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     4321 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     4381 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     4441 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     4501 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct aggccaccat
     4561 ggctgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4621 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4681 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4741 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4801 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4861 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4921 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4981 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5041 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5101 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5161 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5221 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5281 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5341 cgagttcttc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5401 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5461 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5521 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5581 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5641 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     5701 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     5761 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     5821 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     5881 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     5941 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     6001 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc
     6061 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6121 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6181 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6241 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     6301 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     6361 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6421 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     6481 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6541 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     6601 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg
     6661 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6721 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6781 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6841 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6901 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6961 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7441 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7561 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7621 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7681 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7741 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7861 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7921 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7981 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8041 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8101 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8161 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8221 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8281 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8341 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8401 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8461 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8521 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8581 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8641 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8701 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8761 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     8821 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     8881 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     8941 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     9001 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     9061 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     9121 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     9181 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     9241 taaaacgacg gccagtgcca agctg
//
