LOCUS       62056_13810        8916 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 8916)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8916
                     /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        1900..1931
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             1964..3613
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     promoter        3628..4127
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4148..4744
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    4763..5351
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5423..5656
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5734..5868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5895..6030
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6051..6069)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6079..6095)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6237..6692
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6718..6822
                     /label=AmpR promoter
     CDS             6823..7680
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7854..8442
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8730..8751
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8766..8796
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8804..8820
                     /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     8828..8844
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8865..8883
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8911..8916
                     /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 gaacagagtg ttcggtacct agagggtata taatggaagc
     1921 tcgacttcca gcttggcaat ccggtactgt tggtaaagcc accatggaag atgccaaaaa
     1981 cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg gcgagcagct
     2041 gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta ccgacgcaca
     2101 tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc tggcagaagc
     2161 tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg agaatagctt
     2221 gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg ccccagctaa
     2281 cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc ccaccgtcgt
     2341 attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc taccgatcat
     2401 acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa gcatgtacac
     2461 cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc ccgagagctt
     2521 cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg gattgcccaa
     2581 gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc gcgaccccat
     2641 cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg
     2701 cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg tgctcatgta
     2761 ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc aatctgccct
     2821 gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca agtacgacct
     2881 aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc
     2941 cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga cagaaacaac
     3001 cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt
     3061 gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg gtgtgaacca
     3121 gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta acaaccccga
     3181 ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca tcgcctactg
     3241 ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca aatacaaggg
     3301 ctaccaggta gccccagccg aactggagag catcctgctg caacacccca acatcttcga
     3361 cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt
     3421 gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt
     3481 tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact
     3541 gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca agaagggcgg
     3601 caagatcgcc gtgtgagcta gcctaccggg taggggaggc gcttttccca aggcagtctg
     3661 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
     3721 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     3781 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
     3841 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
     3901 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
     3961 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
     4021 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
     4081 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa
     4141 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     4201 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     4261 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     4321 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     4381 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     4441 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     4501 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4561 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4621 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4681 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4741 tgcctgattc tagaaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4801 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4861 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4921 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4981 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5041 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5101 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     5161 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5221 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5281 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5341 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     5401 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     5461 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5521 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     5581 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     5641 tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa
     5701 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     5761 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5821 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     5881 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5941 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     6001 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     6061 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6121 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6181 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     6241 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     6301 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     6361 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     6421 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     6481 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6541 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6601 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6661 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     6721 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6781 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6841 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6901 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6961 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     7021 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     7081 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7141 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7201 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7261 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7321 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7381 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7441 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7501 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7561 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7621 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7681 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7741 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7801 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7861 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7921 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7981 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     8041 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     8101 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8161 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8221 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8281 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8341 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8401 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8461 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     8521 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     8581 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     8641 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     8701 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8761 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     8821 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     8881 agggaacaaa agctggagct gcaagcttaa tgtagt
//
