LOCUS       V042724                 8905 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042724
VERSION     V042724
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..3759
                     /label="GAL3ST4(NM_024637)"
                     /note="GAL3ST4(NM_024637)"
                     /gene="GAL3ST4"
     CDS             3766..3831
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3875..4086
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4099..4367
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4394..4993
                     /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    5000..5588
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5662..5895
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5967..6088
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6107..6242
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6280..6296)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6304..6320
                     /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(6328..6358)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6373..6394
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6682..7270)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7441..8301)
                     /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(8302..8406)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8880..8896
                     /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 catgggccct ctctctcctg ccaggacgct gcggctctgg
     2341 ggacctcgga gcctgggggt ggctctggga gtcttcatga ccattggctt tgcactccag
     2401 ctcttgggag ggcccttcca gaggaggcta cctgggctac agctccgaca gccctcggcc
     2461 ccatccctac gaccagccct tccgtcctgc ccaccccggc agcgactggt gttcctgaag
     2521 acacataaat ccgggagcag ctctgtgctg agcctgcttc accgctatgg ggaccagcac
     2581 gggctgcgct tcgccctccc tgcccgctac cagtttggct acccaaagct cttccaggcc
     2641 tctagggtaa aaggctaccg cccacagggt ggaggcaccc agctcccctt ccacatcctc
     2701 tgtcaccaca tgaggttcaa cctgaaagag gtacttcagg tcatgccttc tgacagcttc
     2761 tttttttcca ttgtccgaga cccagcggct ctggctcgct ctgccttctc ctactataaa
     2821 tccacctcat cagccttccg caagtcacca tctttggctg ccttcctggc caatcctcga
     2881 ggcttctaca ggcctggggc ccgtggggac cactacgctc gcaacttact atggtttgac
     2941 tttggcctgc cctttccccc agagaagagg gccaagagag ggaatattca tccccccaga
     3001 gaccccaacc ccccacagct gcaggtcttg ccttctggtg ctggccctcg agcccaaacc
     3061 ctcaatccca atgccctcat ccatcctgtt tccactgtta ctgatcatcg cagccagata
     3121 tcaagccctg cctctttcga tttggggtct tcatccttca tccagtgggg tctggcatgg
     3181 ctggactctg tctttgacct ggtcatggtg gctgagtact tcgatgagtc attggttctg
     3241 ctggcagatg ccctgtgctg gggtctagat gacgtggtgg gcttcatgca caatgcccag
     3301 gctggacata agcagggcct cagcactgtc agcaacagtg gactgactgc ggaggaccgg
     3361 cagctgactg cacgggcccg agcctggaac aacctggact gggctctcta tgtccacttc
     3421 aaccgcagtc tctgggcacg gatagagaaa tacggccagg gccggctgca gacagctgtg
     3481 gccgagctcc gggctcgccg agaggcccta gcgaaacatt gtctggtagg gggtgaggct
     3541 tctgacccca aatacatcac tgatcgccgg ttccgcccct tccagtttgg gtcagctaag
     3601 gttttgggct atatacttcg gagtggattg agcccccaag accaagagga atgtgagcgc
     3661 ctagctaccc ctgagctcca gtacaaggac aagctggatg ccaagcagtt cccccctacc
     3721 gtctcactgc ccctcaagac ttcaaggcca ctctccccag gatccgacta caaagaccat
     3781 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg
     3841 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     3901 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     3961 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     4021 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     4081 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     4141 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     4201 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     4261 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     4321 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     4381 gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     4441 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     4501 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     4561 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     4621 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     4681 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     4741 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     4801 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     4861 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     4921 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     4981 gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5041 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5101 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5161 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5221 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5281 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     5341 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     5401 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     5461 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     5521 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     5581 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     5641 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc
     5701 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5761 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5821 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5881 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     5941 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     6001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6061 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     6121 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6181 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     6241 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg
     6301 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6361 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6481 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6541 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7081 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7381 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7741 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8401 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     8461 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     8521 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     8581 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     8641 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     8701 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     8761 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     8821 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     8881 taaaacgacg gccagtgcca agctg
//
