LOCUS       V016141                 9554 bp    DNA     circular SYN 25-JAN-2019
DEFINITION  Exported.
ACCESSION   V016141
VERSION     V016141
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9554)
  AUTHORS   Kim J.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-DEC-2018) Biological Sciences, UCSD, 9500 Gilman Dr,
            La Jolla, CA 92093, USA
REFERENCE   2  (bases 1 to 9554)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (18-DEC-2018) Biological Sciences, UCSD, 9500 Gilman Dr, La Jolla,
            CA 92093, USA"
FEATURES             Location/Qualifiers
     source          1..9554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        45..424
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        425..627
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    869..994
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1487..1720
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1905..1949
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             2098..2139
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2247..2364
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    2421..2691
                     /label="tetracycline response element"
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     promoter        2724..2762
                     /label="minimal CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2862..2885
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     promoter        2891..3401
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3423..4019
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4053..4641
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(4644..4660)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             5166..5346
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5378..5602
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5648..6076
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6090..6419
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        6467..6514
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             6533..6904
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     polyA_signal    7037..7170
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7207..7223)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7231..7247)
                     /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)."
     promoter        complement(7255..7285)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7300..7321)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7609..8197)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8371..9228)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9229..9333)
                     /label="AmpR promoter"
ORIGIN
        1 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
       61 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      121 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      181 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      241 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      301 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      361 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      421 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      481 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      541 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      601 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
      661 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      721 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      781 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      841 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      901 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      961 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     1021 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     1081 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     1141 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1201 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1261 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1321 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1381 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1441 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1501 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1561 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1621 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1681 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1741 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     1801 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     1861 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     1921 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     1981 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2041 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2101 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2161 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2221 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2281 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2341 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2401 ttagatctcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
     2461 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     2521 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     2581 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
     2641 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     2701 gtcgagctcg gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag
     2761 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     2821 gaagacaccg ggaccgatcc agcctccgcg gccccgaatt cgattacaaa gacgatgacg
     2881 ataagtagac cggtttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac
     2941 gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg actcgcacat
     3001 tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg
     3061 cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga
     3121 cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag
     3181 caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg
     3241 ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg
     3301 ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag
     3361 tcggctccct cgttgaccga atcaccgacc tctctcccca gggggatcca ccggagctta
     3421 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     3481 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     3541 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     3601 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     3661 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     3721 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     3781 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     3841 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     3901 tggaaacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     3961 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     4021 gagctagcgt taacgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     4081 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     4141 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     4201 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     4261 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     4321 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     4381 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     4441 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     4501 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     4561 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     4621 ccctttgggc cgcctccccg catcgatacc gtcgacctcg agacctagaa aaacatggag
     4681 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
     4741 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
     4801 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     4861 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
     4921 ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt
     4981 gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca
     5041 cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag
     5101 agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact
     5161 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5221 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5281 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     5341 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     5401 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     5461 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     5521 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     5581 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     5641 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5701 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5761 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5821 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5881 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5941 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6001 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6061 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     6121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     6181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     6361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     6421 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     6481 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     6541 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     6601 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     6661 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     6721 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     6781 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     6841 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     6901 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6961 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     7021 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7081 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7141 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     7201 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7261 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7321 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7381 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7441 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7501 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7561 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7621 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7681 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7741 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7801 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7861 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7921 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7981 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8041 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8101 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8161 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8221 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8461 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8641 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8701 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9001 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9241 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9301 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9361 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     9421 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     9481 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     9541 ctgcttaggg ttag
//
