LOCUS       V006415                 7030 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006415
VERSION     V006415
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7030)
  AUTHORS   Siciliano V.
  TITLE     Engineering modular intracellular protein sensor-actuator devices
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7030)
  AUTHORS   Siciliano V.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-APR-2018) Biological Engineering, Massachusetts
            Institute of Technology, 500 Tech Square, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 7030)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7030)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (02-APR-2018) Biological Engineering, Massachusetts Institute of
            Technology, 500 Tech Square, Cambridge, MA 02139, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(222..810)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(984..1841)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1842..1946)
                     /label="AmpR promoter"
     rep_origin      complement(1972..2427)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2569..2585
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2595..2613
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2628..3431
                     /label="HA-L"
                     /note="left homology arm from the adeno-associated virus
                     integration site (AAVS1) within intron 1 of the human
                     PPP1R12C gene"
     misc_feature    3510..3535
                     /label="SA"
                     /note="splice acceptor site"
     protein_bind    4207..4227
                     /label="attB4"
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     regulatory      4273..4427
                     /regulatory_class="promoter"
     protein_bind    4447..4471
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4483..5058
                     /gene="BAX"
                     /label="Apoptosis regulator BAX"
                     /note="Apoptosis regulator BAX from Homo sapiens.
                     Accession#: Q07812"
     protein_bind    complement(5063..5087)
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5242..5297
                     /label="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
     primer_bind     complement(5658..5674)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    5682..5698
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(5706..5736)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    5751..5772
                     /label="CAP binding site"
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    5945..6781
                     /label="HA-R"
                     /note="right homology arm from the adeno-associated virus
                     integration site (AAVS1) within intron 1 of the human
                     PPP1R12C gene"
     promoter        complement(6811..6829)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6850..6866)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6874..6890)
                     /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(6898..6928)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6943..6964)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
       61 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      121 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      181 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      241 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      301 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      361 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      421 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      481 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      541 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      601 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      661 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      721 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      781 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      841 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      901 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      961 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1021 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1081 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1141 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1201 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1261 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1321 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1381 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1441 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1501 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1561 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1621 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1681 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1741 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1801 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1861 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1921 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     1981 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2041 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2101 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2161 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2221 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2281 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2341 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2401 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     2461 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     2521 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2581 ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtacctgc tttctctgac
     2641 cagcattctc tcccctgggc ctgtgccgct ttctgtctgc agcttgtggc ctgggtcacc
     2701 tctacggctg gcccagatcc ttccctgccg cctccttcag gttccgtctt cctccactcc
     2761 ctcttcccct tgctctctgc tgtgttgctg cccaaggatg ctctttccgg agcacttcct
     2821 tctcggcgct gcaccacgtg atgtcctctg agcggatcct ccccgtgtct gggtcctctc
     2881 cgggcatctc tcctccctca cccaacccca tgccgtcttc actcgctggg ttcccttttc
     2941 cttctccttc tggggcctgt gccatctctc gtttcttagg atggccttct ccgacggatg
     3001 tctcccttgc gtcccgcctc cccttcttgt aggcctgcat catcaccgtt tttctggaca
     3061 accccaaagt accccgtctc cctggcttta gccacctctc catcctcttg ctttctttgc
     3121 ctggacaccc cgttctcctg tggattcggg tcacctctca ctcctttcat ttgggcagct
     3181 cccctacccc ccttacctct ctagtctgtg ctagctcttc cagccccctg tcatggcatc
     3241 ttccaggggt ccgagagctc agctagtctt cttcctccaa cccgggcccc tatgtccact
     3301 tcaggacagc atgtttgctg cctccaggga tcctgtgtcc ccgagctggg accaccttat
     3361 attcccaggg ccggttaatg tggctctggt tctgggtact tttatctgtc ccctccaccc
     3421 cacagtgggg caagcttatt accctgttat ccctaaccgg tttaccgagc tcttattggt
     3481 tttcaaactt cattgactgt gccaagcttc tgacctcttc tcttcctccc acagggcctc
     3541 gagacaattg catcgatggt accagtgaaa gtctgcaatg aattcgagtt ggcgcgcctg
     3601 tcattctaaa tctctctttc agcctaaagc tttttccccg tatcccccca ggtgtctgca
     3661 ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc
     3721 ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg
     3781 agccccgggc ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg
     3841 gctttgggag ggggctgtcc ccgtgagctc ccaggcgcgc ctgtcattct aaatctctct
     3901 ttcagcctaa agctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga
     3961 gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac
     4021 gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc
     4081 tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg
     4141 tccccgtgag ctcccaggcg cgccaactcg gtaccgtatc gatgtcgacg ttaacgctag
     4201 tgatatcaac tttgtataga aaagttggct ccgaattggg acagcagaga tccagtttgg
     4261 ttaattaata gacggagtac tgtcctccga gcggagtact gtcctccgac tcgagcggag
     4321 tactgtcctc cgatcggagt actgtcctcc gcgaattccg gagtactgtc ctccgaagac
     4381 gctagcgggg ggctataaaa gggggtgggg gcgttcgtcc tcactctcaa ttcggcccaa
     4441 ttcgacccaa gtttgtacaa aaaagcaggc tgagtcgcca ccatggacgg gtccggggag
     4501 cagcccagag gcggggggcc caccagctct gagcagatca tgaagacagg ggcccttttg
     4561 cttcagggtt tcatccagga tcgagcaggg cgaatggggg gggaggcacc cgagctggcc
     4621 ctggacccgg tgcctcagga tgcgtccacc aagaagctga gcgagtgtct caagcgcatc
     4681 ggggacgaac tggacagtaa catggagctg cagaggatga ttgccgccgt ggacacagac
     4741 tccccccgag aggtcttttt ccgagtggca gctgacatgt tttctgacgg caacttcaac
     4801 tggggccggg ttgtcgccct tttctacttt gccagcaaac tggtgctcaa ggccctgtgc
     4861 accaaggtgc cggaactgat cagaaccatc atgggctgga cattggactt cctccgggag
     4921 cggctgttgg gctggatcca agaccagggt ggttgggacg gcctcctctc ctactttggg
     4981 acgcccacgt ggcagaccgt gaccatcttt gtggcgggag tgctcaccgc ctcgctcacc
     5041 atctggaaga agatgggctg atacccagct ttcttgtaca aagtggtacg cgtgaattca
     5101 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     5161 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     5221 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     5281 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     5341 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     5401 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     5461 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     5521 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     5581 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     5641 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5701 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5761 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5821 cagcggatcg acagtactaa gcttggtgcg tttttatgct tgtagtattg tataatgttt
     5881 ttaagatcct taattaatag ggataacagg gtaatctcga ggtttaaacg tcgacgcggc
     5941 cgctactagg gacaggattg gtgacagaaa agccccatcc ttaggcctcc tccttcctag
     6001 tctcctgata ttgggtctaa cccccacctc ctgttaggca gattccttat ctggtgacac
     6061 acccccattt cctggagcca tctctctcct tgccagaacc tctaaggttt gcttacgatg
     6121 gagccagaga ggatcctggg agggagagct tggcaggggg tgggagggaa gggggggatg
     6181 cgtgacctgc ccggttctca gtggccaccc tgcgctaccc tctcccagaa cctgagctgc
     6241 tctgacgcgg ctgtctggtg cgtttcactg atcctggtgc tgcagcttcc ttacacttcc
     6301 caagaggaga agcagtttgg aaaaacaaaa tcagaataag ttggtcctga gttctaactt
     6361 tggctcttca cctttctagt ccccaattta tattgttcct ccgtgcgtca gttttacctg
     6421 tgagataagg ccagtagcca gccccgtcct ggcagggctg tggtgaggag gggggtgtcc
     6481 gtgtggaaaa ctccctttgt gagaatggtg cgtcctaggt gttcaccagg tcgtggccgc
     6541 ctctactccc tttctctttc tccatccttc tttccttaaa gagtccccag tgctatctgg
     6601 gacatattcc tccgcccaga gcagggtccc gcttccctaa ggccctgctc tgggcttctg
     6661 ggtttgagtc cttggcaagc ccaggagagg cgctcaggct tccctgtccc ccttcctcgt
     6721 ccaccatctc atgcccctgg ctctcctgcc ccttccctac aggggttcct ggctctgctc
     6781 tccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     6841 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6901 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6961 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7021 ttaatgaatc
//
