LOCUS       62056_15600        9156 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 9156)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9156
                     /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        2140..2171
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             2204..3853
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     promoter        3868..4367
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4388..4984
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    5003..5591
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5663..5896
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5974..6108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6135..6270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6291..6309)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6319..6335)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6477..6932
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6958..7062
                     /label=AmpR promoter
     CDS             7063..7920
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8094..8682
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8970..8991
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9006..9036
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9044..9060
                     /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     9068..9084
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9105..9123
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9151..9156
                     /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 ggctcgagag gaacatgtcc caacatgttg agctctagag gaagaagact
     1921 gggcatgtct gggcaaggaa catgtcccaa catgttgagc tctagaggaa gaagactggg
     1981 catgtctggg caaggaacat gtcccaacat gttgagctct agaggaagaa gactgggcat
     2041 gtctgggcaa ggaacatgtc ccaacatgtt gagctctaga ggaagaagac tgggcatgtc
     2101 tgggcaagga acatgtccca acatgttgag ctcggtacct agagggtata taatggaagc
     2161 tcgacttcca gcttggcaat ccggtactgt tggtaaagcc accatggaag atgccaaaaa
     2221 cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg gcgagcagct
     2281 gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta ccgacgcaca
     2341 tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc tggcagaagc
     2401 tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg agaatagctt
     2461 gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg ccccagctaa
     2521 cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc ccaccgtcgt
     2581 attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc taccgatcat
     2641 acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa gcatgtacac
     2701 cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc ccgagagctt
     2761 cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg gattgcccaa
     2821 gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc gcgaccccat
     2881 cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg
     2941 cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg tgctcatgta
     3001 ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc aatctgccct
     3061 gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca agtacgacct
     3121 aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc
     3181 cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga cagaaacaac
     3241 cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt
     3301 gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg gtgtgaacca
     3361 gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta acaaccccga
     3421 ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca tcgcctactg
     3481 ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca aatacaaggg
     3541 ctaccaggta gccccagccg aactggagag catcctgctg caacacccca acatcttcga
     3601 cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt
     3661 gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt
     3721 tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact
     3781 gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca agaagggcgg
     3841 caagatcgcc gtgtgagcta gcctaccggg taggggaggc gcttttccca aggcagtctg
     3901 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
     3961 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     4021 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
     4081 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
     4141 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
     4201 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
     4261 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
     4321 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa
     4381 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     4441 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     4501 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     4561 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     4621 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     4681 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     4741 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4801 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4861 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4921 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4981 tgcctgattc tagaaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     5041 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5101 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5161 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5221 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5281 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5341 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     5401 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5461 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5521 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5581 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     5641 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     5701 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5761 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     5821 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     5881 tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa
     5941 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     6001 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6061 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     6121 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6181 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     6241 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     6301 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6361 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6421 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     6481 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     6541 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     6601 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     6661 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     6721 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6781 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6841 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6901 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     6961 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     7021 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     7081 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     7141 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     7201 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     7261 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     7321 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7381 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7441 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7501 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7561 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7621 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7681 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7741 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7801 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7861 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7921 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7981 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     8041 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     8101 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     8161 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     8221 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     8281 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     8341 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8401 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8461 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8521 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8581 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8641 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8701 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     8761 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     8821 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     8881 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     8941 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     9001 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     9061 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     9121 agggaacaaa agctggagct gcaagcttaa tgtagt
//
