LOCUS       V029595                 9595 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029595
VERSION     V029595
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..4449
                     /label="IKBKE(NM_014002)"
                     /note="IKBKE(NM_014002)"
                     /gene="IKBKE"
     CDS             4456..4521
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4565..4776
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4789..5057
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5084..5683
                     /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    5690..6278
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6352..6585
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6657..6778
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6797..6932
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6970..6986)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6994..7010
                     /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(7018..7048)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7063..7084
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7372..7960)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8131..8991)
                     /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(8992..9096)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9570..9586
                     /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 catgcagagc acagccaatt acctgtggca cacagatgac
     2341 ctgctggggc agggggccac tgccagtgtg tacaaggccc gcaacaagaa atccggagag
     2401 ctggttgctg tgaaggtctt caacactacc agctacctgc ggccccgcga ggtgcaggtg
     2461 agggagtttg aggtcctgcg gaagctgaac caccagaaca ttgtcaagct ctttgcggtg
     2521 gaggagacgg gcggaagccg gcagaaggta ctggtgatgg agtactgctc cagtgggagc
     2581 ctgctgagtg tgctggagag ccctgagaat gcctttgggc tgcctgagga tgagttcctg
     2641 gtggtgctgc gctgtgtggt ggccggcatg aaccacctgc gggagaacgg cattgtgcat
     2701 cgcgacatca agccggggaa catcatgcgc ctcgtagggg aggaggggca gagcatctac
     2761 aagctgacag acttcggcgc tgcccgggag ctggatgatg atgagaagtt cgtctcggtc
     2821 tatgggactg aggagtacct gcatcccgac atgtatgagc gggcggtgct tcgaaagccc
     2881 cagcaaaaag cgttcggggt gactgtggat ctctggagca ttggagtgac cttgtaccat
     2941 gcagccactg gcagcctgcc cttcatcccc tttggtgggc cacggcggaa caaggagatc
     3001 atgtaccgga tcaccacgga gaagccggct ggggccattg caggtgccca gaggcgggag
     3061 aacgggcccc tggagtggag ctacaccctc cccatcacct gccagctgtc actggggctg
     3121 cagagccagc tggtgcccat cctggccaac atcctggagg tggagcaggc caagtgctgg
     3181 ggcttcgacc agttctttgc ggagaccagt gacatcctgc agcgagttgt cgtccatgtc
     3241 ttctccctgt cccaggcagt cctgcaccac atctatatcc atgcccacaa cacgatagcc
     3301 attttccagg aggccgtgca caagcagacc agtgtggccc cccgacacca ggagtacctc
     3361 tttgagggtc acctctgtgt cctcgagccc agcgtctcag cacagcacat cgcccacacg
     3421 acggcaagca gccccctgac cctcttcagc acagccatcc ctaaggggct ggccttcagg
     3481 gaccctgctc tggacgtccc caagttcgtc cccaaagtgg acctgcaggc ggattacaac
     3541 actgccaagg gcgtgttggg cgccggctac caggccctgc ggctggcacg ggccctgctg
     3601 gatgggcagg agctaatgtt tcgggggctg cactgggtca tggaggtgct ccaggccaca
     3661 tgcagacgga ctctggaagt ggcaaggaca tccctcctct acctcagcag cagcctggga
     3721 actgagaggt tcagcagcgt ggctggaacg cctgagatcc aggaactgaa ggcggctgca
     3781 gaactgaggt ccaggctgcg gactctagcg gaggtcctct ccagatgctc ccaaaatatc
     3841 acggagaccc aggagagcct gagcagcctg aaccgggagc tggtgaagag ccgggatcag
     3901 gtacatgagg acagaagcat ccagcagatt cagtgctgtt tggacaagat gaacttcatc
     3961 tacaaacagt tcaagaagtc taggatgagg ccagggcttg gctacaacga ggagcagatt
     4021 cacaagctgg ataaggtgaa tttcagtcat ttagccaaaa gactcctgca ggtgttccag
     4081 gaggagtgcg tgcagaagta tcaagcgtcc ttagtcacac acggcaagag gatgagggtg
     4141 gtgcacgaga ccaggaacca cctgcgcctg gttggctgtt ctgtggctgc ctgtaacaca
     4201 gaagcccagg gggtccagga gagtctcagc aagctcctgg aagagctatc tcaccagctc
     4261 cttcaggacc gagcaaaggg ggctcaggcc tcgccgcctc ccatagctcc ttaccccagc
     4321 cctacacgaa aggacctgct tctccacatg caagagctct gcgaggggat gaagctgctg
     4381 gcatctgacc tcctggacaa caaccgcatc atcgaacggc taaatagagt cccagcacct
     4441 cctgatgtcg gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     4501 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     4561 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     4621 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     4681 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     4741 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     4801 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     4861 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     4921 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     4981 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     5041 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc
     5101 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     5161 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     5221 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     5281 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     5341 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     5401 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     5461 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5521 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     5581 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5641 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct
     5701 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5761 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5821 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5881 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5941 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     6001 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     6061 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     6121 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     6181 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     6241 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga
     6301 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     6361 taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag
     6421 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6481 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6541 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     6601 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     6661 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6721 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6781 atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6841 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6901 ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc
     6961 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7021 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7081 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7141 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7201 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7261 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7321 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7381 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7441 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7501 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7561 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7621 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7681 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7741 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7801 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7861 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7921 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7981 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8041 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8101 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8161 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8221 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8281 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8341 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8401 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8461 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8521 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8581 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8641 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8701 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8761 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8821 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8881 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8941 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9001 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9061 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9121 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     9181 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     9241 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     9301 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     9361 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     9421 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     9481 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     9541 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
