LOCUS       V016031                 9360 bp    DNA     circular SYN 28-JUL-2021
DEFINITION  Exported.
ACCESSION   V016031
VERSION     V016031
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9360)
  AUTHORS   Liu L, Limsakul P, Wang Y.
  TITLE     Integration of FRET and Sequencing to Engineer Kinase Biosensors
            from Mammalian Cell Libraries
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9360)
  AUTHORS   Liu L, Limsakul P, Wang Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUL-2021) Department of Bioengineering, Institute of
            Engineering in Medicine, University of California, San Diego, 9500
            Gilman Drive 0435, La Jolla, CA 92093-0435, USA
REFERENCE   3  (bases 1 to 9360)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (10-JUL-2021) Department of Bioengineering, Institute of Engineering
            in Medicine, University of California, San Diego, 9500 Gilman Drive
            0435, La Jolla, CA 92093-0435, USA"
FEATURES             Location/Qualifiers
     source          1..9360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             117..750
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    797..922
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1415..1648
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1833..1877
                     /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             2026..2067
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2164..2281
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2409..3587
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             3610..4293
                     /label="ECFP"
                     /note="enhanced CFP"
     misc_feature    4681..5394
                     /note="YPet; receptor"
     misc_feature    5476..6062
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             6082..6678
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             6884..7117
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7274..7498
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        7594..7698
                     /label="AmpR promoter"
     CDS             7699..8556
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      8730..9318
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 cggaccgcca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
       61 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttgga
      121 agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct accacacaca
      181 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
      241 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
      301 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
      361 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
      421 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
      481 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
      541 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
      601 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      661 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      721 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      781 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      841 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      901 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
      961 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1021 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1081 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1141 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1201 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1261 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1321 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1381 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1441 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1501 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1561 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1621 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1681 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     1741 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     1801 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     1861 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     1921 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     1981 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     2041 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     2101 gatccattcg attagtgaac ggatctcgac ggtatcgcca caaatggcag tattcatcca
     2161 caattttaaa agaaaggggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     2221 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     2281 tcgggtttat tacagggaca gcagagatcc actttggtcg ataagctttg caaagatgga
     2341 taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag
     2401 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     2461 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
     2521 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     2581 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     2641 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
     2701 cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
     2761 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
     2821 aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
     2881 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
     2941 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
     3001 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
     3061 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
     3121 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
     3181 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
     3241 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
     3301 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     3361 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     3421 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     3481 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     3541 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgagga attgatcctt
     3601 cgaactagta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3661 gagctggacg gcgacgtaaa cggccacagg ttcagcgtgt ccggcgaggg cgagggcgat
     3721 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3781 tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcagccg ctaccccgac
     3841 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3901 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3961 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4021 ctggggcaca agctggagta caactatatc agccacaacg tctatatcac cgccgacaag
     4081 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcagcgtg
     4141 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4201 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaagga gaagcgcgat
     4261 cacatggtcc tgctggagtt cgtgaccgcc gcccgcatgc attggtattt tgggaagatc
     4321 actcgtcggg agtccgagcg gctgctgctc aaccccgaaa acccccgggg aaccttcttg
     4381 gtccgggaga gcgagacgac aaaaggtgcc tattgcctct ccgtttctga ctttgacaac
     4441 gccaaggggc tcaatgtgaa gcactacaag atccgcaagc tggacagcgg cggcttctac
     4501 atcacctcac gcacacagtt cagcagcctg cagcagctgg tggcctacta ctccaaacat
     4561 gctgatggct tgtgccaccg cctgaccaac gtctgcgggt ctacatctgg atctgggaag
     4621 ccgggttctg gtgagggttc ttcaagggag tacgcctgca tcagtggtga gctggagctc
     4681 tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat
     4741 gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac gtacggtaaa
     4801 ttgaccttaa aattactctg tactactggt aaattgccag ttccatggcc aaccttagtc
     4861 actactttag gttatggtgt tcaatgtttt gctagatacc cagatcatat gaaacaacat
     4921 gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa
     4981 gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat
     5041 agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg
     5101 gaatacaact ataactctca caatgtttac atcactgctg acaaacaaaa gaatggtatc
     5161 aaagctaact tcaaaattag acacaacatt gaagatggtg gtgttcaatt agctgaccat
     5221 tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta
     5281 tcctatcaat ctgccttatt caaagatcca aacgaaaaga gagaccacat ggtcttgtta
     5341 gaatttttga ctgctgctgg tattaccgag ggtatgaatg aattgtacaa ataggaattc
     5401 gaagcttgag ctcgagatct gagtccggta gcgctagttc gaaggatccg catgcatcta
     5461 gggcggccaa ttccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt
     5521 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     5581 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     5641 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     5701 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     5761 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     5821 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     5881 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     5941 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     6001 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag cttgccacaa
     6061 cccacaagga gacgaccttc catgaccgag tacaagccca cggtgcgcct cgccacccgc
     6121 gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     6181 cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     6241 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     6301 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     6361 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     6421 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     6481 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     6541 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     6601 ctcggcttca ccgtcaccgc cgacgtcgag tgcccgaagg accgcgcgac ctggtgcatg
     6661 acccgcaagc ccggtgcctg ataataggcg gccgctcgag acctagaaaa acatggagca
     6721 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     6781 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     6841 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     6901 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     6961 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7021 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7081 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     7141 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattat
     7201 aatagattta gcaggaattg aactaggagt ggagcacaca ggcaaagttc tagagctcgc
     7261 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     7321 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     7381 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     7441 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     7501 tctgaggcgg aaagaaccag ctgggggcgc gcccctcgag gccgccatgg tcatagctgt
     7561 ttgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     7621 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7681 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7741 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7801 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7861 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7921 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     7981 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     8041 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     8101 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     8161 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     8221 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     8281 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     8341 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     8401 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     8461 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     8521 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     8581 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8641 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8701 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8761 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8821 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     8881 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     8941 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     9001 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     9061 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     9121 cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     9181 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     9241 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     9301 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
//
