LOCUS       V043868                 9282 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043868
VERSION     V043868
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3301
                     /label="HOXA1(NM_005522)"
                     /note="HOXA1(NM_005522)"
                     /gene="HOXA1"
     CDS             3308..3397
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3441..3652
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3665..3933
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3962..3971
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3992..4654
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4724..4777
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4778..5377
                     /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    5378..5965
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6039..6272
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6344..6465
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6484..6619
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6657..6673)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6681..6697
                     /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(6705..6735)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6750..6771
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7059..7647)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7818..8678)
                     /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(8679..8783)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9257..9273
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgg acaatgcaag aatgaactcc ttcctggaat accccatact
     2341 tagcagtggc gactcgggga cctgctcagc ccgagcctac ccctcggacc ataggattac
     2401 aactttccag tcgtgcgcgg tcagcgccaa cagttgcggc ggcgacgacc gcttcctagt
     2461 gggcaggggg gtgcagatcg gttcgcccca ccaccaccac caccaccacc atcgccaccc
     2521 ccagccggct acctaccaga cttccgggaa cctgggggtg tcctactccc actcaagttg
     2581 tggtccaagc tatggctcac agaacttcag tgcgccttac agcccctacg cgttaaatca
     2641 ggaagcagac gtaagtggtg ggtaccccca gtgcgctccc gctgtttact ctggaaatct
     2701 ctcatctccc atggtccagc atcaccacca ccaccagggt tatgctgggg gcgcggtggg
     2761 ctcgcctcaa tacattcacc actcatatgg acaggagcac cagagcctgg ccctggctac
     2821 gtataataac tccttgtccc ctctccacgc cagccaccaa gaagcctgtc gctcccccgc
     2881 atcggagaca tcttctccag cgcagacttt tgactggatg aaagtcaaaa gaaaccctcc
     2941 caaaacaggg aaagttggag agtacggcta cctgggtcaa cccaacgcgg tgcgcaccaa
     3001 cttcactacc aagcagctca cggaactgga gaaggagttc cacttcaaca agtacctgac
     3061 gcgcgcccgc agggtggaga tcgctgcatc cctgcagctc aacgagaccc aagtgaagat
     3121 ctggttccag aaccgccgaa tgaagcaaaa gaaacgtgag aaggagggtc tcttgcccat
     3181 ctctccggcc accccgccag gaaacgacga gaaggccgag gaatcctcag agaagtccag
     3241 ctcttcgccc tgcgttcctt ccccggggtc ttctacctca gacactctga ctacctccca
     3301 cggatccgaa caaaaactaa tatcagagga agatttggaa cagaaactaa tcagtgaaga
     3361 agatttagag caaaagttaa tttccgagga ggacttatag cggccgcgaa ggatctgcga
     3421 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     3481 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag
     3541 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     3601 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct
     3661 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     3721 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     3781 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     3841 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt
     3901 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga
     3961 cgccaccatg gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac
     4021 cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccaagca
     4081 gggccgcatg accaacaaga tgaagagcac caaaggcgcc ctgaccttca gcccctacct
     4141 gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga
     4201 gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta
     4261 cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg gccgcgtgat
     4321 cggcgacttc aaggtggtgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa
     4381 gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata acgtgctggt
     4441 gggcagcttc gcccgcacct tcagcctgcg cgacggcggc tactacagct tcgtggtgga
     4501 cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat
     4561 gttcgccttc cgccgcgtgg aggagctgca cagcaacacc gagctgggca tcgtggagta
     4621 ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc cgcgctcagt cgtccaattc
     4681 tgccgtggac ggcaccgccg gacccggctc caccggatct cgcgagggca gaggaagtct
     4741 tctaacatgc ggtgacgtgg aggagaatcc cggccctatg accgagtaca agcccacggt
     4801 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     4861 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     4921 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     4981 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     5041 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     5101 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     5161 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     5221 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     5281 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5341 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat caacctctgg attacaaaat
     5401 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5461 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5521 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5581 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5641 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5701 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5761 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     5821 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5881 cggcctgctg ccggctctgc ggcctcttcc gcgtctccgc cttcgccctc agacgagtcg
     5941 gatctccctt tggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc
     6001 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     6061 acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     6121 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6181 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6241 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     6301 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     6361 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6421 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     6481 gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6541 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6601 ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca
     6661 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6721 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6781 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8521 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     8821 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     8881 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     8941 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     9001 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     9061 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     9121 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     9181 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     9241 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
