LOCUS       V044064                 9055 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044064
VERSION     V044064
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..3915
                     /label="Mgat3(NM_010795)"
                     /note="Mgat3(NM_010795)"
                     /gene="Mgat3"
     CDS             3916..3981
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4025..4236
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4249..4517
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4544..5143
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5150..5738
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5812..6045
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6117..6238
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6257..6392
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6430..6446)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6454..6470
                     /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(6478..6508)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6523..6544
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6832..7420)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7591..8451)
                     /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(8452..8556)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9030..9046
                     /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 catgaagatg agacgctaca agctctttct catgttctgt
     2341 atggctggcc tgtgcctcat atccttcctg cacttcttta agaccttatc ctatgtcacc
     2401 ttcccgagag aactggcctc cctcagccct aacctcgtat ccagcttctt ctggaacaat
     2461 gcccctgtca ctccccaggc cagtccggag ccgggtggcc ccgacctatt gcggacaccc
     2521 ctctactccc actctcccct gctccagcca ctgtccccga gcaaggccac agaggaactg
     2581 caccgggtgg acttcgtgtt gccggaggac accacggagt attttgtgcg caccaaagct
     2641 ggtggtgtgt gcttcaaacc aggtaccagg atgctggaga aaccttcgcc agggcggaca
     2701 gaggagaagc ccgaagtgtc tgagggctcc tcagcccggg gacctgctcg gaggcccatg
     2761 aggcacgtgt tgagtacgcg ggagcgcctg ggcagccggg gcactaggcg caagtgggtt
     2821 gagtgtgtgt gcctgccagg ctggcacggg cccagttgcg gggtgcccac ggtggtgcag
     2881 tattccaacc tgcccaccaa ggaacgcctg gtacccaggg aggtaccgag gcgggttatc
     2941 aacgccatca acatcaacca cgagttcgac ctgctggatg tgcgtttcca tgagctggga
     3001 gatgttgtgg acgccttcgt ggtctgtgaa tctaatttca ccgcctacgg ggagcctcgg
     3061 ccgctcaagt tccgagagat gctgaccaat ggcaccttcg agtacatccg ccacaaggtg
     3121 ctctatgtct tcctggacca tttcccacct ggtggccgtc aggacggctg gattgcggat
     3181 gactacctgc gcaccttcct cacccaggat ggcgtctccc gcctgcgcaa cctgcggccc
     3241 gatgacgtct ttatcatcga cgatgcggac gagatccctg cgcgtgatgg tgtgctgttc
     3301 ctcaaactct acgatggctg gacagagccc ttcgccttcc acatgcggaa gtccctgtat
     3361 ggtttcttct ggaagcagcc gggcacactg gaggtggtgt caggctgcac catggacatg
     3421 ctgcaggccg tgtatgggct ggatggcatc cgcctgcgcc gccgccagta ctacaccatg
     3481 cccaacttcc ggcagtatga gaaccgcacc ggccacatcc tagtgcagtg gtctctcggc
     3541 agccccctgc acttcgcggg ctggcattgc tcctggtgct tcacacccga gggcatctac
     3601 tttaaactcg tgtcagccca gaatggcgac ttcccccgct ggggtgacta tgaggacaag
     3661 agggacctca attacatccg cagcttgatc cgcactgggg gatggttcga cggaacgcag
     3721 caggagtacc ctcctgcgga ccccagtgag cacatgtatg ctcctaaata cctgctcaag
     3781 aactatgacc agttccgcta cttgctggaa aatccctacc gggagcccaa gagcactgaa
     3841 gagggtgggc gccggaacca gggctcagat ggaaggccat ctgctgtcag gggcaagttg
     3901 gatacagtgg agggcgacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3961 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     4021 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     4081 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     4141 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     4201 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     4261 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     4321 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     4381 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     4441 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     4501 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc
     4561 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     4621 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     4681 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4741 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4801 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4861 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4921 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4981 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     5041 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5101 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct
     5161 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5221 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5281 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5341 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5401 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5461 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5521 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     5581 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5641 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     5701 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga
     5761 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     5821 taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag
     5881 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5941 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6001 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     6061 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     6121 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6181 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6241 atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6301 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6361 ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc
     6421 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6481 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6541 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6601 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6661 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6721 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6781 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6841 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6901 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6961 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7021 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7081 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7141 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7201 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7261 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7321 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7381 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7441 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7501 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7561 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7621 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7681 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7741 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7801 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7861 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7921 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7981 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8041 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8101 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8161 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8221 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8281 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8341 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8401 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8461 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8521 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8581 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     8641 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     8701 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     8761 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     8821 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     8881 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     8941 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     9001 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
