LOCUS       V044138                 9620 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044138
VERSION     V044138
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"
     regulatory      2296..2305
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2305..2346
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             2353..4539
                     /label="TRDN(NM_006073)"
                     /note="TRDN(NM_006073)"
                     /gene="TRDN"
     promoter        4590..4801
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4814..5082
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5109..5708
                     /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    5715..6303
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6377..6610
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6682..6803
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6822..6957
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6995..7011)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7019..7035
                     /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(7043..7073)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7088..7109
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7397..7985)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8156..9016)
                     /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(9017..9121)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9595..9611
                     /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 catgggtaag cctatcccta accctctcct cggtctcgat
     2341 tctacactcg agatgactga gatcactgct gaaggaaatg catctacaac cacaactgtg
     2401 atagacagca aaaatggatc tgtgcccaaa tcccccggaa aagtgctgaa gaggacagtc
     2461 acagaagaca tagtgacgac gttcagctcc cctgcagcct ggcttctggt cattgccctg
     2521 ataatcacgt ggtcagctgt tgccatcgtt atgtttgatt tagtggatta caaaaacttt
     2581 tcagcaagct ctattgccaa gattggctca gatcctttaa aactggtacg tgatgctatg
     2641 gaggaaacca cggactggat ctatggcttc ttttctttgt tatctgacat catctcatct
     2701 gaagatgaag aagatgatga tggtgacgaa gatactgata aaggagaaat agatgagcct
     2761 cccttgagaa aaaaagaaat acacaaagat aagactgaaa aacaagagaa acctgaaagg
     2821 aaaatacaaa ctaaagttac acacaaagaa aaagaaaaag gaaaagaaaa agtaagagaa
     2881 aaagaaaaac ctgaaaagaa agcaactcac aaggaaaaaa ttgagaaaaa agaaaaacca
     2941 gaaacaaaga cactggcgaa agaacagaag aaagctaaga ctgcagaaaa gagtgaagaa
     3001 aagactaaaa aggaagtgaa aggtggaaaa caggagaaag tgaagcaaac agctgcaaaa
     3061 gtaaaagaag tacagaaaac accatcaaaa cccaaagaaa aggaggacaa agagaaagca
     3121 gctgtgtcaa agcatgaaca gaaagatcag tatgcattct gtcgatatat gattgacata
     3181 tttgtccatg gggatttaaa accaggacaa agcccagcca ttccacctcc cttaccgaca
     3241 gaacaagctt ccagacccac tccggcatca cctgcccttg aagaaaaaga aggggaaaag
     3301 aagaaggctg agaagaaagt tacttctgaa acaaaaaaga aagaaaaaga agatatcaaa
     3361 aagaaaagtg agaaggaaac tgccattgat gtggaaaaaa aagagccggg aaaagcttct
     3421 gaaaccaaac aagggactgt aaaaattgca gcacaagcag cagctaagaa ggatgaaaag
     3481 aaggaagatt ccaagaaaac aaaaaaacct gcagaagtag aacaacccaa gggaaaaaaa
     3541 caggaaaaga aagaaaaaca tgtggaacca gcaaagtcac caaagaaaga acactcagtt
     3601 ccaagtgaca aacaagtaaa agcaaaaact gaacgagcca aagaggagat tggtgcggtt
     3661 tcaattaaaa aagctgtacc tggaaagaag gaagagaaaa caaccaagac agtggagcaa
     3721 gaaattagaa aagaaaaatc tgggaagact tcttcaattc tgaaagataa agaacctatt
     3781 aaagggaaag aagagaaagt tccagcttcc ctaaaggaaa aagaacctga aactaaaaaa
     3841 gatgaaaaga tgtccaaagc aggcaaagaa gtcaagccta aacctccaca actacaagga
     3901 aaaaaggaag agaagccaga gccccaaatt aaaaaagaag caaaaccagc tatatctgaa
     3961 aaagtgcaaa tacacaaaca agacatagtg aaaccagaaa agactgtttc tcatggtaaa
     4021 ccagaagaaa aagttctcaa gcaggtaaaa gctgtcacaa tagaaaaaac agccaagccc
     4081 aaaccaacaa aaaaagctga acatcgagaa agagaacctc catctataaa aacagacaaa
     4141 ccaaaaccaa ctccaaaagg aacatcagaa gtcacagaat caggaaagaa gaaaactgaa
     4201 atatctgaaa aagaaagtaa agaaaaagca gacatgaagc atcttagaga agaaaaagtc
     4261 tcaacaagaa aagaaagtct tcaattacac aatgtgacaa aagcagaaaa acctgcaaga
     4321 gtatcaaaag atgttgaaga tgtaccagct tcaaagaaag ctaaagaagg aactgaagat
     4381 gtgtctccca caaagcagaa aagtcccatc agtttcttcc agtgtgtcta cttggatggg
     4441 tacaatggct atggatttca gtttcctttc actcctgcag accgccctgg agagagctct
     4501 ggtcaagcaa attctccagg acagaagcaa caaggacagt aaggatccgc ggccgcgaag
     4561 gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     4621 gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa
     4681 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     4741 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     4801 gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg
     4861 ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc
     4921 gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc
     4981 ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact
     5041 ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc
     5101 tacgctagat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc
     5161 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     5221 acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     5281 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     5341 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     5401 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     5461 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     5521 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     5581 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     5641 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     5701 gtgcctgagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     5761 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     5821 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     5881 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     5941 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     6001 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     6061 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     6121 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     6181 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     6241 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     6301 cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6361 aaagaaaagg ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt
     6421 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6481 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6541 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6601 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     6661 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     6721 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6781 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     6841 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6901 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6961 acttttgcag agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt
     7021 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     7081 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     7141 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     7201 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     7261 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7321 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7381 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7441 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7501 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7561 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7621 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7681 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7741 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7801 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7861 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7921 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7981 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     8041 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     8101 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     8161 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8221 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8281 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8341 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8401 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8461 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8521 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8581 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8641 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8701 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8761 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8821 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8881 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8941 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     9001 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     9061 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     9121 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     9181 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     9241 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     9301 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     9361 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     9421 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
     9481 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     9541 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     9601 cgacggccag tgccaagctg
//
