LOCUS       62056_15620        8522 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 8522)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        1879..2236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2287..3219
                     /codon_start=1
                     /label=hRluc
                     /note="Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        3234..3733
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3754..4350
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    4369..4957
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5029..5262
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5340..5474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5657..5675)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5685..5701)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5843..6298
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6324..6428
                     /label=AmpR promoter
     CDS             6429..7286
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7460..8048
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8336..8357
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8372..8402
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8410..8426
                     /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     8434..8450
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8471..8489
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8517..8522
                     /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 ggctcgagct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     1921 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1981 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2041 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2101 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2161 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2221 cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaacc aaaggctgga
     2281 gccaccatgg cttccaaggt gtacgacccc gagcaacgca aacgcatgat cactgggcct
     2341 cagtggtggg ctcgctgcaa gcaaatgaac gtgctggact ccttcatcaa ctactatgat
     2401 tccgagaagc acgccgagaa cgccgtgatt tttctgcatg gtaacgctgc ctccagctac
     2461 ctgtggaggc acgtcgtgcc tcacatcgag cccgtggcta gatgcatcat ccctgatctg
     2521 atcggaatgg gtaagtccgg caagagcggg aatggctcat atcgcctcct ggatcactac
     2581 aagtacctca ccgcttggtt cgagctgctg aaccttccaa agaaaatcat ctttgtgggc
     2641 cacgactggg gggcttgtct ggcctttcac tactcctacg agcaccaaga caagatcaag
     2701 gccatcgtcc atgctgagag tgtcgtggac gtgatcgagt cctgggacga gtggcctgac
     2761 atcgaggagg atatcgccct gatcaagagc gaagagggcg agaaaatggt gcttgagaat
     2821 aacttcttcg tcgagaccat gctcccaagc aagatcatgc ggaaactgga gcctgaggag
     2881 ttcgctgcct acctggagcc attcaaggag aagggcgagg ttagacggcc taccctctcc
     2941 tggcctcgcg agatccctct cgttaaggga ggcaagcccg acgtcgtcca gattgtccgc
     3001 aactacaacg cctaccttcg ggccagcgac gatctgccta agatgttcat cgagtccgac
     3061 cctgggttct tttccaacgc tattgtcgag ggagctaaga agttccctaa caccgagttc
     3121 gtgaaggtga agggcctcca cttcagccag gaggacgctc cagatgaaat gggtaagtac
     3181 atcaagagct tcgtggagcg cgtgctgaag aacgagcagt aagctagcct accgggtagg
     3241 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
     3301 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
     3361 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
     3421 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
     3481 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
     3541 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
     3601 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
     3661 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
     3721 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc
     3781 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     3841 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     3901 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     3961 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4021 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4081 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4141 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     4201 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4261 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     4321 acctggtgca tgacccgcaa gcccggtgcc tgattctaga aagtcgacaa tcaacctctg
     4381 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4441 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4501 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4561 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4621 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4681 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4741 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     4801 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4861 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4921 cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt
     4981 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     5041 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     5101 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     5161 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     5221 gatccctcag acccttttag tcagtgtgga aaatctctag cagtacgtat agtagttcat
     5281 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     5341 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5401 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5461 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     5521 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5581 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5641 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     5701 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5761 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5821 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5881 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5941 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6001 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6061 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6121 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6181 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6241 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     6301 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6361 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6421 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6481 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6541 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6601 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6661 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6721 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6781 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6841 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6901 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6961 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7021 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7081 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7141 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7201 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7261 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7321 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7381 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7441 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7501 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7561 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7621 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7681 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7741 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7801 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7861 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7921 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7981 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8041 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8101 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     8161 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     8221 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     8281 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     8341 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     8401 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     8461 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta
     8521 gt
//
