LOCUS       62056_17215        9003 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 9003)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9003
                     /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             3341..4054
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     protein_bind    complement(4520..4544)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4546..4587
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4629..5217
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5289..5522
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5600..5734
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5761..5896
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5917..5935)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5945..5961)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6103..6558
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6584..6688
                     /label=AmpR promoter
     CDS             6689..7546
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7720..8308
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8596..8617
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8632..8662
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8670..8686
                     /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     8694..8710
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8731..8749
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8777..9003
                     /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 agcaggctac
     3241 catgtccgtc ctgacgccgc tgctgctgcg gggcttgaca ggctcggccc ggcggctccc
     3301 agtgccgcgc gccaagatcc attcgttgcc gccgggatcc gtgagcaagg gcgaggagct
     3361 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     3421 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     3481 ctccaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     3541 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     3601 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     3661 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     3721 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaactg
     3781 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     3841 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     3901 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcacct gctccgccct
     3961 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     4021 cgggatcact ctcggcatgg acgagctgta caagactagt ggtggttcag gtggtggtgg
     4081 ttcaggtggt ggtggttcag gtggaggagg atcaggagga ggaggatcag gaggaggagg
     4141 atcaggagga gaattcgctc aagagtttgt gaactgcaaa atccagcctg ggaaggtggt
     4201 tgtgttcatc aagcccacct gcccgtactg caggagggcc caagagatcc tcagtcaatt
     4261 gcccatcaaa caagggcttc tggaatttgt cgatatcaca gccaccaacc acactaacga
     4321 gattcaagat tatttgcaac agctcacggg agcaagaacg gtgcctcgag tctttattgg
     4381 taaagattgt ataggcggat gcagtgatct agtctctttg caacagagtg gggaactgct
     4441 gacgcggcta aagcagattg gagctctgca gggtacctct agaggatctt tgtgaaggaa
     4501 ccttacttct gtggtgtgaa cccagctttc ttgtacaaag tggttggtaa gcctatccct
     4561 aaccctctcc tcggtctcga ttctacgtag taatgagcta gcgctaaccg gtggcgcgtt
     4621 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     4681 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     4741 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     4801 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     4861 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     4921 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4981 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     5041 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     5101 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     5161 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
     5221 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     5281 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     5341 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     5401 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     5461 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     5521 cagtacgtat agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     5581 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     5641 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5701 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     5761 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5821 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5881 ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc
     5941 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     6001 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     6061 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     6121 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     6181 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     6241 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     6301 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     6361 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6421 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6481 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     6541 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7861 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     8041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     8101 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     8161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     8221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     8281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     8401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     8461 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     8521 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     8581 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     8641 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     8701 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     8761 ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg
     8821 atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa
     8881 gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga
     8941 cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata
     9001 aac
//
