LOCUS       V028562                 9349 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028562
VERSION     V028562
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..4002
                     /label="Slc47a1(NM_026183)"
                     /note="Slc47a1(NM_026183)"
                     /gene="Slc47a1"
     CDS             4009..4074
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4118..4329
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4342..4610
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4643..5437
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5444..6032
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6106..6339
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6411..6532
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6551..6686
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6724..6740)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6748..6764
                     /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(6772..6802)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6817..6838
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7126..7714)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7885..8745)
                     /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(8746..8850)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9324..9340
                     /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 catggaacgc acggaggagt ccgctcccgg gccgggaggc
     2341 gcggacgctg cgtccgagcg ccgtgggcta cgctgcctgc tgctgcccgg cttcctagag
     2401 gagctgcggg cactcttggt cctggcgggt cctgcgttct tggcccagct gatgatgttc
     2461 ctaatcagtt tcataagctc agtgttctgt ggccacttgg gcaagctgga gttggatgca
     2521 gtcacactgg caattgcggt tatcaatgtc acaggcattt cagtgggaca tggcttgtct
     2581 tctgcttgtg acacgctcat ctcccagaca tacgggagcc agaacttaaa gcacgtaggg
     2641 gttatcctgc aaagggggac actcatcctg ctcctctgct gctttccctg ctgggcactc
     2701 ttcatcaaca ccgagcaaat tctgctgctc ttcagacagg acccggatgt atccaggctc
     2761 acccagactt acgtcatgat cttcatccca gcgcttcctg cagcttttct ttatacgtta
     2821 caagttaaat acttgctcaa ccagggcatc gttctgcctc agattatgac gggcatcgca
     2881 gctaaccttg tcaacgccct ggccaactat gtgtttctct atcacctgca tcttggggtg
     2941 atgggttcag cattggccaa caccatctcc cagtttgccc tggccatttt cctgttcctc
     3001 tacatcctct ggagaagact acaccaagcc acctggggag ggtggtcctg ggagtgtctg
     3061 caggactggg cctccttcct gcgactggct atccccagca tgctcatgct gtgcatagag
     3121 tggtgggcct atgaggttgg cagcttcctg agtggtatcc ttggcatggt ggagttggga
     3181 gcccagtcca tcacctatga attggctatc attgtataca tgatcccttc aggcttcagt
     3241 gtggctgcca acgtccgggt gggaaatgca ctgggcgcag gaaacattga ccaggccaag
     3301 aagtcctcag ctatttcctt gatagtcaca gagctctttg ctgtgacctt ctgcgtcctg
     3361 ctgctaggct gtaaggatct cgtgggctac attttcacta ccgatcggga tatcgtggcc
     3421 ttggtggctc aagtgatccc catttatgct gtgtcccacc tctttgaagg tcttgcttgt
     3481 acctgtggtg gtatcctgag gggcactggg aaccagaagg ttggagccat cgttaatgcc
     3541 atcgggtatt atgtcattgg cctccccatc gggatcgcgc tgatgtttgc tgctaagctg
     3601 ggagtgatcg gcctgtggtc aggcatcatc atctgcacta cctgtcagac cacgtgcttc
     3661 ctggctttta ttgctcggct caattggaaa cgcgcctgtc aacaggctca agtacatgcc
     3721 aatttgaagg taaacgtggc cctgaattcc gctgtctctc acgagcctgc tcacccagtg
     3781 tgtcctgaga gccatggaga aattatgatg acggatcttg aaaaaaaaga cgagactcag
     3841 ttggaccagc cgatgaacca gcaacaagct ttgcctatcc gtccaaagga cagcaataaa
     3901 ctgtctggga aacagttggc cttacggaga ggactcctgc tcctaggggt agtcttagtc
     3961 ttggtgggtg ggattttagt gagagtctat atcagaattg aaggatccga ctacaaagac
     4021 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg
     4081 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     4141 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc
     4201 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     4261 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     4321 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc
     4381 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc
     4441 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg
     4501 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt
     4561 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga
     4621 ccggcgccta cgctaggcca ccatggctga acaagatgga ttgcacgcag gttctccggc
     4681 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4741 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4801 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4861 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4921 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4981 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5041 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5101 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5161 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5221 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5281 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5341 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5401 catcgccttc tatcgccttc ttgacgagtt cttctgagtc gacaatcaac ctctggatta
     5461 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5521 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5581 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5641 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5701 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5761 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5821 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     5881 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5941 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6001 gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca atgacttaca
     6061 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     6121 actcccaacg aaaataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     6181 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6241 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6301 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     6361 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     6421 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6481 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6541 ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6601 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6661 gtagtgagga ggcttttttg gaggcctaga cttttgcaga gacggcccaa attcgtaatc
     6721 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6781 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6841 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6901 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6961 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7021 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7081 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7561 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7801 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7861 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7981 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8041 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8401 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8761 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8821 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8881 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     8941 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     9001 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     9061 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     9121 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     9181 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     9241 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     9301 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
