LOCUS       V032023                 9279 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032023
VERSION     V032023
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3361
                     /label="HNRNPA2B1(NM_031243)"
                     /note="HNRNPA2B1(NM_031243)"
                     /gene="HNRNPA2B1"
     CDS             3368..3433
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3477..3688
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3701..3969
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4004..4711
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             4721..4774
                     /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             4775..5374
                     /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    5375..5962
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6036..6269
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6341..6462
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6481..6616
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6654..6670)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6678..6694
                     /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(6702..6732)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6747..6768
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7056..7644)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7815..8675)
                     /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(8676..8780)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9254..9270
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatggagaa aactttagaa actgttcctt tggagaggaa
     2341 aaagagagaa aaggaacagt tccgtaagct ctttattggt ggcttaagct ttgaaaccac
     2401 agaagaaagt ttgaggaact actacgaaca atggggaaag cttacagact gtgtggtaat
     2461 gagggatcct gcaagcaaaa gatcaagagg atttggtttt gtaacttttt catccatggc
     2521 tgaggttgat gctgccatgg ctgcaagacc tcattcaatt gatgggagag tagttgagcc
     2581 aaaacgtgct gtagcaagag aggaatctgg aaaaccaggg gctcatgtaa ctgtgaagaa
     2641 gctgtttgtt ggcggaatta aagaagatac tgaggaacat caccttagag attactttga
     2701 ggaatatgga aaaattgata ccattgagat aattactgat aggcagtctg gaaagaaaag
     2761 aggctttggc tttgttactt ttgatgacca tgatcctgtg gataaaatcg tattgcagaa
     2821 ataccatacc atcaatggtc ataatgcaga agtaagaaag gctttgtcta gacaagaaat
     2881 gcaggaagtt cagagttcta ggagtggaag aggaggcaac tttggctttg gggattcacg
     2941 tggtggcggt ggaaatttcg gaccaggacc aggaagtaac tttagaggag gatctgatgg
     3001 atatggcagt ggacgtggat ttggggatgg ctataatggg tatggaggag gacctggagg
     3061 tggcaatttt ggaggtagcc ccggttatgg aggaggaaga ggaggatatg gtggtggagg
     3121 acctggatat ggcaaccagg gtgggggcta cggaggtggt tatgacaact atggaggagg
     3181 aaattatgga agtggaaatt acaatgattt tggaaattat aaccagcaac cttctaacta
     3241 cggtccaatg aagagtggaa actttggtgg tagcaggaac atggggggac catatggtgg
     3301 aggaaactat ggtccaggag gcagtggagg aagtgggggt tatggtggga ggagccgata
     3361 cggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     3421 tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     3481 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     3541 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     3601 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     3661 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     3721 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     3781 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     3841 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     3901 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     3961 ttacagatcc aagctgtgac cggcgcctac gctagacgcc accatggtga gcaagggcga
     4021 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     4081 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     4141 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     4201 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     4261 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     4321 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     4381 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     4441 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     4501 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     4561 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     4621 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     4681 ctccaccggc ggcatggacg agctgtacaa gggatctcgc gagggcagag gaagtcttct
     4741 aacatgcggt gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg
     4801 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     4861 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     4921 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4981 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5041 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5101 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5161 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5221 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5281 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5341 cacctggtgc atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg
     5401 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5461 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5521 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5581 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5641 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5701 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5761 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5821 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5881 cctgctgccg gctctgcggc ctcttccgcg tctccgcctt cgccctcaga cgagtcggat
     5941 ctccctttgg ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt
     6001 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     6061 aaaataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     6121 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     6181 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     6241 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     6301 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     6361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6421 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     6481 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6541 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     6601 ggcttttttg gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag
     6661 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     6721 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     6781 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6841 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6901 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6961 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7021 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7081 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7141 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7201 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7261 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7321 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7381 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7441 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7501 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7561 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7621 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7681 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7741 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7801 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7861 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7921 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7981 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8041 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8101 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8161 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8221 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8281 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8341 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8401 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8461 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8521 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8581 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8641 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8701 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8761 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     8821 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     8881 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     8941 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     9001 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     9061 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     9121 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     9181 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     9241 cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
