LOCUS       V028142                 8729 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028142
VERSION     V028142
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..3487
                     /label="B3GNT2(NM_001319075)"
                     /note="B3GNT2(NM_001319075)"
                     /gene="B3GNT2"
     promoter        3532..3743
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3756..4024
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4053..4062
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4083..4745
                     /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"
     misc_feature    4824..5412
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5486..5719
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5791..5912
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5931..6066
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6104..6120)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6128..6144
                     /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(6152..6182)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6197..6218
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6506..7094)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7265..8125)
                     /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(8126..8230)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8704..8720
                     /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 gccaccatga gtgttggacg tcgaagaata aagttgttgg gtatcctgat
     2341 gatggcaaat gtcttcattt attttattat ggaagtctcc aaaagcagta gccaagaaaa
     2401 aaatggaaaa ggggaagtaa taatacccaa agagaagttc tggaagatat ctacccctcc
     2461 cgaggcatac tggaaccgag agcaagagaa gctgaaccgg cagtacaacc ccatcctgag
     2521 catgctgacc aaccagacgg gggaggcggg caggctctcc aatataagcc atctgaacta
     2581 ctgcgaacct gacctgaggg tcacgtcggt ggttacgggt tttaacaact tgccggacag
     2641 atttaaagac tttctgctgt atttgagatg ccgcaattat tcactgctta tagatcagcc
     2701 ggataagtgt gcaaagaaac ctttcttgtt gctggcgatt aagtccctca ctccacattt
     2761 tgccagaagg caagcaatcc gggaatcctg gggccaagaa agcaacgcag ggaaccaaac
     2821 ggtggtgcga gtcttcctgc tgggccagac acccccagag gacaaccacc ccgacctttc
     2881 agatatgctg aaatttgaga gtgagaagca ccaagacatt cttatgtgga actacagaga
     2941 cactttcttc aacttgtctc tgaaggaagt gctgtttctc aggtgggtaa gtacttcctg
     3001 cccagacact gagtttgttt tcaagggcga tgacgatgtt tttgtgaaca cccatcacat
     3061 cctgaattac ttgaatagtt tatccaagac caaagccaaa gatctcttca taggtgatgt
     3121 gatccacaat gctggacctc atcgggataa gaagctgaag tactacatcc cagaagttgt
     3181 ttactctggc ctctacccac cctatgcagg gggagggggg ttcctctact ccggccacct
     3241 ggccctgagg ctgtaccata tcactgacca ggtccatctc taccccattg atgacgttta
     3301 tactggaatg tgccttcaga aactcggcct cgttccagag aaacacaaag gcttcaggac
     3361 atttgatatc gaggagaaaa acaaaaataa catctgctcc tatgtagatc tgatgttagt
     3421 acatagtaga aaacctcaag agatgattga tatttggtct cagttgcaga gtgctcattt
     3481 aaaatgctaa gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc
     3541 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct
     3601 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     3661 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     3721 acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg
     3781 cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg
     3841 cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac
     3901 cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt
     3961 gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca
     4021 gatccaagct gtgaccggcg cctacgctag acgccaccat ggagagcgac gagagcggcc
     4081 tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg gagttcgagc
     4141 tggtgggcgg cggagagggc acccccaagc agggccgcat gaccaacaag atgaagagca
     4201 ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc tacggcttct
     4261 accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc atcaacaacg
     4321 gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg cacgtgagct
     4381 tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg ggcaccggct
     4441 tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc accgtggagc
     4501 acctgcaccc catgggcgat aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc
     4561 gcgacggcgg ctactacagc ttcgtggtgg acagccacat gcacttcaag agcgccatcc
     4621 accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc
     4681 acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc cccatcgcct
     4741 tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct
     4801 ccaccggatc tcgctaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     4861 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4921 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4981 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     5041 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     5101 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     5161 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     5221 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     5281 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     5341 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     5401 ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5461 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat
     5521 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5581 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5641 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5701 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     5761 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     5821 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5881 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     5941 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6001 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     6061 gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg
     6121 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6181 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6241 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6301 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6361 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6421 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6481 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6541 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6601 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6661 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6721 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6781 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6841 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6901 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6961 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7021 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7081 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7141 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7201 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7261 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7321 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7381 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7441 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7501 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7561 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7621 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7681 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7741 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7801 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7861 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7921 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7981 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8041 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8101 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8221 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     8281 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     8341 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     8401 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     8461 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     8521 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     8581 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     8641 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     8701 gttgtaaaac gacggccagt gccaagctg
//
