LOCUS       62056_15605        9033 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 9033)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             222..402
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    449..574
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1067..1300
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1485..1529
                     /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"
     misc_feature    1713..1830
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2017..2048
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             2081..3730
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     promoter        3745..4244
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4265..4861
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    4880..5468
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5540..5773
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5851..5985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6012..6147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6168..6186)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6196..6212)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6354..6809
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6835..6939
                     /label=AmpR promoter
     CDS             6940..7797
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7971..8559
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8847..8868
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8883..8913
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8921..8937
                     /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     8945..8961
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8982..9000
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9028..9033
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac
       61 aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct
      121 tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt
      181 gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc
      241 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      301 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      361 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      421 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      481 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      541 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      601 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      661 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      721 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
      781 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
      841 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
      901 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
      961 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1021 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1081 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1141 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1201 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1261 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1321 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     1381 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     1441 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     1501 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     1561 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     1621 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     1681 acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg gattgggggg
     1741 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     1801 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     1861 ctttggcgcc ggctcgaggc atgcctgcag gtcgacaggg gaccaggaca aaggtcacgt
     1921 tcgggagtcg acaggggacc aggacaaagg tcacgttcgg gagtcgacag gggaccagga
     1981 caaaggtcac gttcgggagt cgactctaga ggtacctaga gggtatataa tggaagctcg
     2041 acttccagct tggcaatccg gtactgttgg taaagccacc atggaagatg ccaaaaacat
     2101 taagaagggc ccagcgccat tctacccact cgaagacggg accgccggcg agcagctgca
     2161 caaagccatg aagcgctacg ccctggtgcc cggcaccatc gcctttaccg acgcacatat
     2221 cgaggtggac attacctacg ccgagtactt cgagatgagc gttcggctgg cagaagctat
     2281 gaagcgctat gggctgaata caaaccatcg gatcgtggtg tgcagcgaga atagcttgca
     2341 gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc cagctaacga
     2401 catctacaac gagcgcgagc tgctgaacag catgggcatc agccagccca ccgtcgtatt
     2461 cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa aagaagctac cgatcataca
     2521 aaagatcatc atcatggata gcaagaccga ctaccagggc ttccaaagca tgtacacctt
     2581 cgtgacttcc catttgccac ccggcttcaa cgagtacgac ttcgtgcccg agagcttcga
     2641 ccgggacaaa accatcgccc tgatcatgaa cagtagtggc agtaccggat tgcccaaggg
     2701 cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg accccatctt
     2761 cggcaaccag atcatccccg acaccgctat cctcagcgtg gtgccatttc accacggctt
     2821 cggcatgttc accacgctgg gctacttgat ctgcggcttt cgggtcgtgc tcatgtaccg
     2881 cttcgaggag gagctattct tgcgcagctt gcaagactat aagattcaat ctgccctgct
     2941 ggtgcccaca ctatttagct tcttcgctaa gagcactctc atcgacaagt acgacctaag
     3001 caacttgcac gagatcgcca gcggcggggc gccgctcagc aaggaggtag gtgaggccgt
     3061 ggccaaacgc ttccacctac caggcatccg ccagggctac ggcctgacag aaacaaccag
     3121 cgccattctg atcacccccg aaggggacga caagcctggc gcagtaggca aggtggtgcc
     3181 cttcttcgag gctaaggtgg tggacttgga caccggtaag acactgggtg tgaaccagcg
     3241 cggcgagctg tgcgtccgtg gccccatgat catgagcggc tacgttaaca accccgaggc
     3301 tacaaacgct ctcatcgaca aggacggctg gctgcacagc ggcgacatcg cctactggga
     3361 cgaggacgag cacttcttca tcgtggaccg gctgaagagc ctgatcaaat acaagggcta
     3421 ccaggtagcc ccagccgaac tggagagcat cctgctgcaa caccccaaca tcttcgacgc
     3481 cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg cccgccgcag tcgtcgtgct
     3541 ggaacacggt aaaaccatga ccgagaagga gatcgtggac tatgtggcca gccaggttac
     3601 aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac gaggtgccta aaggactgac
     3661 cggcaagttg gacgcccgca agatccgcga gattctcatt aaggccaaga agggcggcaa
     3721 gatcgccgtg tgagctagcc taccgggtag gggaggcgct tttcccaagg cagtctggag
     3781 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     3841 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     3901 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     3961 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
     4021 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     4081 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg
     4141 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca
     4201 cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct
     4261 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     4321 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     4381 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     4441 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     4501 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     4561 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     4621 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     4681 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     4741 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     4801 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     4861 ctgattctag aaagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     4921 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     4981 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5041 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5101 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5161 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     5221 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     5281 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     5341 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     5401 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     5461 ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     5521 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     5581 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     5641 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     5701 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     5761 aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt
     5821 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     5881 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5941 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     6001 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6061 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6121 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     6181 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     6241 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     6301 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     6361 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6421 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6481 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6541 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6601 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6661 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6721 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6781 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     6841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6901 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6961 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7021 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7081 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7141 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7201 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7261 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7321 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7381 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7441 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7501 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7561 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7621 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7681 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7741 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7801 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     8041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     8101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     8161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     8221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     8461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     8581 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     8641 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     8701 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     8761 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     8821 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     8881 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     8941 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     9001 gaacaaaagc tggagctgca agcttaatgt agt
//
