LOCUS       V029505                 9805 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029505
VERSION     V029505
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..4707
                     /label="H6PD(NM_001282587)"
                     /note="H6PD(NM_001282587)"
                     /gene="H6PD"
     CDS             4714..4731
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        4775..4986
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4999..5267
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5294..5893
                     /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    5900..6488
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6562..6795
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6867..6988
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7007..7142
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7180..7196)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7204..7220
                     /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(7228..7258)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7273..7294
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7582..8170)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8341..9201)
                     /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(9202..9306)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9780..9796
                     /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 catgttagca gagcctttta actggcaccc aggcatgtgg
     2341 aatatgctca tagtggcgat gtgcttggcc cttctgggct gcctgcaagc ccaggagctc
     2401 cagggacatg tctccataat cctgctggga gcaactgggg acctggctaa gaagtactta
     2461 tggcagggac tgttccagct gtacctggat gaagcgggga ggggtcacag ttttagcttc
     2521 catggagctg ctctgacagc ccccaagcag ggtcaagagc tcatggccaa ggccctggaa
     2581 tccctctcct gccccaagga catggcaccc agtcactgtg cagagcacaa ggatcagttc
     2641 ctgcagctga gccagtaccg ccaactgaag acggccgagg actatcaggc cctgaacaag
     2701 gacatcgagg cacagctcca gcacgcaggc ctccgggagg ctggcaggat cttctacttc
     2761 tcagtgccac ccttcgccta tgaagacatt gcccgcaaca tcaacagtag ctgccggcca
     2821 ggcccgggcg cctggctgcg ggttgtcctt gagaaaccct ttggccatga ccacttctca
     2881 gcccagcagc tggccacaga actcgggacc tttttccagg aggaggagat gtaccgggtg
     2941 gaccattact taggcaagca ggctgtggcg cagatcctgc ctttccgaga ccagaaccgc
     3001 aaggctttgg acggcctctg gaaccggcac catgtggagc gggtggagat catcatgaaa
     3061 gagaccgtgg atgctgaagg ccgcaccagc ttctatgagg agtacggtgt cattcgcgac
     3121 gtcctccaga accatctgac ggaggtcctc accctcgtgg ccatggagct gccccacaat
     3181 gtcagcagtg cggaggctgt gctgcggcac aagcttcagg tcttccaggc gctgcggggc
     3241 ctgcagaggg gcagtgccgt cgtgggccag taccagtctt acagtgagca ggtgcgcaga
     3301 gagctgcaga agccagacag cttccacagc ctgacgccga ccttcgcagc cgtcctagtg
     3361 cacattgaca accttcgctg ggagggcgtg cctttcatcc tgatgtctgg caaagccttg
     3421 gacgagagag tgggctacgc tcggatcttg ttcaagaacc aggcctgctg tgtgcagagc
     3481 gaaaagcact gggccgcggc gcagagccag tgcctgcccc ggcagctcgt cttccacatc
     3541 ggccatggcg acctgggcag ccctgccgtg ctggtcagca ggaacctgtt caggccctcc
     3601 ctgccctcca gctggaagga aatggaggga ccacctgggc tccgcctttt cggcagccct
     3661 ctgtccgatt actacgccta cagccctgtg cgggagcggg acgcccactc cgtcctctta
     3721 tcccatatct tccatggccg gaagaatttc ttcatcacca cagagaactt gctggcctcc
     3781 tggaacttct ggacccctct gctggagagc ctggcccata aggccccacg cctctaccct
     3841 ggaggagctg agaatggccg tctgttggac tttgagttca gtagcggccg gttgttcttt
     3901 tcccagcagc agccggagca gctggtgcca gggccagggc cggccccaat gcccagtgac
     3961 ttccaggtcc tcagggccaa gtaccgagag agcccgctgg tctccgcctg gtccgaggag
     4021 ctgatctcta agctggctaa tgacatcgag gccaccgctg tgcgagccgt gcggcgcttt
     4081 ggccagttcc acctggcact gtcggggggc tcgagccccg tggccctgtt ccagcagctg
     4141 gccacggcgc actatggctt cccctgggcc cacacgcacc tgtggctggt tgacgagcgc
     4201 tgcgtcccac tctcagaccc ggagtccaac ttccagggcc tgcaggccca cctgctgcag
     4261 cacgtccgga tcccctacta caacatccac cccatgcctg tgcacctgca gcagcggctc
     4321 tgcgccgagg aggaccaggg cgcccagatc tatgccaggg agatctcagc cctggtggcc
     4381 aacagcagct tcgacctggt gctgctgggc atgggtgccg acgggcacac agcctccctc
     4441 ttcccacagt cacccactgg cctggatggc gagcagctgg tcgtgctgac cacgagcccc
     4501 tcccagccac accgccgcat gagccttagc ctgcctctca tcaaccgcgc caagaaggtg
     4561 gcagtcctgg tcatgggcag gatgaagcgt gagatcacca cgctggtgag ccgggtgggc
     4621 catgagccca agaagtggcc catctcgggt gtcctgccgc actccggcca gctggtgtgg
     4681 tacatggact acgacgcctt cctgggagga tcccatcacc atcaccatca ctagcggccg
     4741 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     4801 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     4861 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     4921 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     4981 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     5041 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     5101 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     5161 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     5221 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     5281 gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     5341 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     5401 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     5461 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     5521 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     5581 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     5641 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     5701 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     5761 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     5821 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     5881 gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5941 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     6001 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     6061 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     6121 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     6181 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6241 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     6301 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6361 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6421 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6481 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     6541 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc
     6601 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6661 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6721 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6781 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     6841 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     6901 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6961 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     7021 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     7081 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     7141 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg
     7201 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7261 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7321 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7381 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7441 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7501 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7561 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7621 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7681 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7741 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7801 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7861 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7921 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7981 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     8041 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8101 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8161 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8221 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8281 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8401 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8461 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8521 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8581 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8641 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8701 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8761 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8821 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8881 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8941 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     9001 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     9061 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9121 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9181 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9301 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     9361 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     9421 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     9481 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     9541 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     9601 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     9661 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     9721 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     9781 taaaacgacg gccagtgcca agctg
//
