LOCUS       pCDH-CMV-Fluc-EF        9837 bp DNA     circular SYN 20-OCT-2024
DEFINITION  Exported.
ACCESSION   V013705
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9837)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9837)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..233
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             234..414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1076..1309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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             1686..1727
                     /label=Protein Tat
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    1798..1915
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2005..2208
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2297..3946
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     promoter        3996..4207
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             4220..4488
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4544..5209
                     /label=CopGFP
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             5279..5332
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5333..5929
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    5933..6520
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6594..6827
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6899..7033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      7039..7174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7212..7228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7236..7252)
                     /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(7260..7290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7305..7326)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7614..8202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8376..9233)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9234..9338)
                     /label=AmpR promoter
     primer_bind     9812..9828
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta
     2341 tccgctagag gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct
     2401 ggttcctgga acaattgctt ttacagatgc acatatcgag gtgaacatca cgtacgcgga
     2461 atacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa
     2521 tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc
     2581 gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct
     2641 caacagtatg aacatttcgc agcctaccgt agtgtttgtt tccaaaaagg ggttgcaaaa
     2701 aattttgaac gtgcaaaaaa aattaccaat aatccagaaa attattatca tggattctaa
     2761 aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg
     2821 ttttaatgaa tacgattttg taccagagtc ctttgatcgt gacaaaacaa ttgcactgat
     2881 aatgaactcc tctggatcta ctgggttacc taagggtgtg gcccttccgc atagaactgc
     2941 ctgcgtcaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac
     3001 tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta cactcggata
     3061 tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttttacg
     3121 atcccttcag gattacaaaa ttcaaagtgc gttgctagta ccaaccctat tttcattctt
     3181 cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg
     3241 gggcgcacct ctttcgaaag aagtcgggga agcggttgca aaacgcttcc atcttccagg
     3301 gatacgacaa ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg
     3361 ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga
     3421 tctggatacc gggaaaacgc tgggcgttaa tcagagaggc gaattatgtg tcagaggacc
     3481 tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga
     3541 tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact tcttcatagt
     3601 tgaccgcttg aagtctttaa ttaaatacaa aggataccag gtggcccccg ctgaattgga
     3661 gtcgatattg ttacaacacc ccaacatctt cgacgcgggc gtggcaggtc ttcccgacga
     3721 tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga
     3781 aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg
     3841 agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat
     3901 cagagagatc ctcataaagg ccaagaaggg cggaaagtcc aaattgtgag gatccgcggc
     3961 cgcaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     4021 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     4081 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     4141 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     4201 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     4261 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     4321 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     4381 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     4441 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc
     4501 ggcgcctacg ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat
     4561 cgagtgccgc atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga
     4621 gggcaccccc aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac
     4681 cttcagcccc tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta
     4741 ccccagcggc tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac
     4801 ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga
     4861 ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt
     4921 gatcttcacc gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg
     4981 cgataacgtg ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta
     5041 cagcttcgtg gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca
     5101 gaacgggggc cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct
     5161 gggcatcgtg gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc
     5221 tcagtcgtcc aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga
     5281 gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga
     5341 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     5401 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     5461 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5521 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5581 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5641 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5701 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5761 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5821 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5881 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc
     5941 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     6001 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     6061 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     6121 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     6181 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     6241 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     6301 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     6361 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6421 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc tccgccttcg
     6481 ccctcagacg agtcggatct ccctttggcc gcctccccgc ctggtacctt taagaccaat
     6541 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     6601 gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt
     6661 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6721 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6781 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     6841 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     6901 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6961 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     7021 tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc
     7081 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     7141 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat
     7201 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     7261 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     7321 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7381 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7441 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7501 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7561 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7621 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7681 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7741 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7801 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7861 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7921 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7981 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     8041 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     8101 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     8161 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     8221 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     8281 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     8341 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8401 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8461 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8521 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8581 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8641 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8701 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8761 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8821 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8881 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8941 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     9001 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     9061 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     9121 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     9181 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     9241 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     9301 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9361 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     9421 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     9481 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     9541 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     9601 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     9661 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     9721 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     9781 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
