LOCUS       62056_17220        9175 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9175)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /codon_start=1
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
                     /translation="KNEQELLELDKWASL"
     promoter        1483..1983
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             1995..2390
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    2450..2567
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2684..2987
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2988..3191
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    3214..3238
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3376..4089
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     protein_bind    complement(4692..4716)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4718..4759
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4801..5389
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5461..5694
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5772..5906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5933..6068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6089..6107)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6117..6133)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6275..6730
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6756..6860
                     /label=AmpR promoter
     CDS             6861..7718
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7892..8480
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8768..8789
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8804..8834
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8842..8858
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8866..8882
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8903..8921
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8949..9175
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccttgcgcct tttccaaggc
     1501 agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc
     1561 gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc
     1621 gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc
     1681 ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc
     1741 gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat
     1801 agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg
     1861 gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag
     1921 cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc
     1981 cagggggtac caccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
     2041 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
     2101 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
     2161 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
     2221 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
     2281 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
     2341 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc ctgcagctgc
     2401 agtagtaaga attctagatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa
     2461 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     2521 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     2581 gggacagcag agatccactt tggcgccggc tcgagggggc ccggggttat taatagtaat
     2641 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     2701 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     2761 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     2821 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     2881 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     2941 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     3001 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     3061 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     3121 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     3181 taagcagagc tctctggcta actgtcggga tcaacaagtt tgtacaaaaa agcaggcttt
     3241 aaaggaacca attcagtcga ctggatctgc ggccgcatgt ccgtcctgac gccgctgctg
     3301 ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc cgcgcgccaa gatccattcg
     3361 ttgccgccgg gatccgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3421 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3481 gatgccacct acggcaagct gaccctgaag ttcatctcca ccaccggcaa gctgcccgtg
     3541 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     3601 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3661 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3721 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3781 atcctggggc acaagctgga gtacaactac aactgccaca acgtctatat catggccgac
     3841 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3901 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3961 cccgacaacc actacctgag cacctgctcc gccctgagca aagaccccaa cgagaagcgc
     4021 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     4081 ctgtacaaga ctagtggtgg ttcaggtggt ggtggttcag gtggtggtgg ttcaggtgga
     4141 ggaggatcag gaggaggagg atcaggagga ggaggatcag gaggagaatt cgatatctca
     4201 gaattctata agctagcacc tgttgacaag aaaggccaac cattcccctt cgaccaatta
     4261 aagggaaaag tggtgcttat cgttaatgtt gcctccaaat gtggattcac tcctcaatac
     4321 aaagaactag aggccttgta caaacgttat aaggacgaag gatttaccat catcgggttc
     4381 ccatgcaacc agtttggcca ccaagaacct ggctctgatg aagaaattgc ccagttctgc
     4441 caactgaact atggcgtgac tttccccatt atgaaaaaaa ttgacgttaa tggtggcaat
     4501 gaggaccctg tttacaagtt tttgaagagc caaaaatccg gtatgttggg cttgagaggt
     4561 atcaaatgga attttgaaaa attcttagtc gataaaaagg gtaaagtgta cgaaagatac
     4621 tcttcactaa ccaaaccttc ttcgttgtcc gaaaccatcg aagaactttt gaaagaggtg
     4681 gaatagtcta gacccagctt tcttgtacaa agtggttggt aagcctatcc ctaaccctct
     4741 cctcggtctc gattctacgt agtaatgagc tagcgctaac cggtggcgcg ttaagtcgac
     4801 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4861 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4921 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4981 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5041 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5101 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5161 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     5221 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5281 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     5341 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     5401 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     5461 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     5521 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     5581 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     5641 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtacgt
     5701 atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     5761 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5821 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5881 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     5941 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6001 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6061 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     6121 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     6181 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     6241 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     6301 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     6361 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6421 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     6481 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     6541 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     6601 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     6661 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     6721 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6781 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6841 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6901 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6961 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     7021 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     7081 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     7141 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     7201 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     7261 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     7321 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     7381 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     7441 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7501 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7561 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7621 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7681 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7741 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7801 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7861 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7921 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7981 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     8041 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     8101 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     8161 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     8221 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     8281 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     8341 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8401 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8461 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8521 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8581 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     8641 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8701 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8761 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     8821 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     8881 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
     8941 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
     9001 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
     9061 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
     9121 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaac
//
