LOCUS       40924_1849        7209 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector LR_UAS-EYFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7209)
  AUTHORS   Siciliano V.
  TITLE     Engineering modular intracellular protein sensor-actuator devices
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7209)
  AUTHORS   Siciliano V.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-APR-2018) Biological Engineering, Massachusetts 
            Institute of Technology, 500 Tech Square, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 7209)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7209)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-APR-2018) Biological Engineering, Massachusetts Institute of 
            Technology, 500 Tech Square, Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(160..1017)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1018..1122)
                     /label=AmpR promoter
     rep_origin      complement(1148..1603)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1745..1761
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1771..1789
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1804..2607
                     /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    2686..2711
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    3383..3403
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     regulatory      3448..3600
                     /regulatory_class="promoter"
     protein_bind    3623..3647
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3674..4390
                     /label=EYFP
                     /note="enhanced YFP"
     protein_bind    complement(4418..4442)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4597..4652
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5013..5029)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5037..5053
                     /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(5061..5091)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5106..5127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    5300..6136
                     /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(6166..6184)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6205..6221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6229..6245)
                     /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(6253..6283)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6298..6319)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6607..7195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
       61 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
      121 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
      181 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
      241 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
      301 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
      361 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
      421 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
      481 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
      541 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
      601 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
      661 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
      721 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
      781 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
      841 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
      901 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
      961 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     1021 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1081 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     1141 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1201 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     1261 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     1321 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     1381 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     1441 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     1501 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     1561 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     1621 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     1681 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     1741 cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt
     1801 acctgctttc tctgaccagc attctctccc ctgggcctgt gccgctttct gtctgcagct
     1861 tgtggcctgg gtcacctcta cggctggccc agatccttcc ctgccgcctc cttcaggttc
     1921 cgtcttcctc cactccctct tccccttgct ctctgctgtg ttgctgccca aggatgctct
     1981 ttccggagca cttccttctc ggcgctgcac cacgtgatgt cctctgagcg gatcctcccc
     2041 gtgtctgggt cctctccggg catctctcct ccctcaccca accccatgcc gtcttcactc
     2101 gctgggttcc cttttccttc tccttctggg gcctgtgcca tctctcgttt cttaggatgg
     2161 ccttctccga cggatgtctc ccttgcgtcc cgcctcccct tcttgtaggc ctgcatcatc
     2221 accgtttttc tggacaaccc caaagtaccc cgtctccctg gctttagcca cctctccatc
     2281 ctcttgcttt ctttgcctgg acaccccgtt ctcctgtgga ttcgggtcac ctctcactcc
     2341 tttcatttgg gcagctcccc tacccccctt acctctctag tctgtgctag ctcttccagc
     2401 cccctgtcat ggcatcttcc aggggtccga gagctcagct agtcttcttc ctccaacccg
     2461 ggcccctatg tccacttcag gacagcatgt ttgctgcctc cagggatcct gtgtccccga
     2521 gctgggacca ccttatattc ccagggccgg ttaatgtggc tctggttctg ggtactttta
     2581 tctgtcccct ccaccccaca gtggggcaag cttattaccc tgttatccct aaccggttta
     2641 ccgagctctt attggttttc aaacttcatt gactgtgcca agcttctgac ctcttctctt
     2701 cctcccacag ggcctcgaga caattgcatc gatggtacca gtgaaagtct gcaatgaatt
     2761 cgagttggcg cgcctgtcat tctaaatctc tctttcagcc taaagctttt tccccgtatc
     2821 cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt
     2881 gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag
     2941 cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt
     3001 aattacatcc ctgggggctt tgggaggggg ctgtccccgt gagctcccag gcgcgcctgt
     3061 cattctaaat ctctctttca gcctaaagct ttttccccgt atccccccag gtgtctgcag
     3121 gctcaaagag cagcgagaag cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc
     3181 cgggctgtcc ccgcacgctg ccggctcggg gatgcggggg gagcgccgga ccggagcgga
     3241 gccccgggcg gctcgctgct gccccctagc gggggaggga cgtaattaca tccctggggg
     3301 ctttgggggg gggctgtccc cgtgagctcc caggcgcgcc aactcggtac cgtatcgatg
     3361 tcgacgttaa cgctagtgat atcaactttg tatagaaaag ttggctccga attgggacag
     3421 cagagatcca gtttggttaa ttaatagacg gagtactgtc ctccgagcgg agtactgtcc
     3481 tccgactcga gcggagtact gtcctccgat cggagtactg tcctccgcga attccggagt
     3541 actgtcctcc gaagacgcta gcggggggct ataaaagggg gtgggggcgt tcgtcctcac
     3601 tctcaattcg gcccaattcg acccaagttt gtacaaaaaa gcaggcttaa ccggttctgg
     3661 acatatggcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3721 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3781 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     3841 gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc
     3901 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3961 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     4021 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     4081 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     4141 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     4201 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     4261 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     4321 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     4381 gctgtacaag taagcggccg ctcctgcatt aagctttacc cagctttctt gtacaaagtg
     4441 gtacgcgtga attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     4501 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     4561 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     4621 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     4681 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     4741 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     4801 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     4861 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     4921 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     4981 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5041 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5101 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5161 ggaaacctgt cgtgccagcg gatcgacagt actaagcttg gtgcgttttt atgcttgtag
     5221 tattgtataa tgtttttaag atccttaatt aatagggata acagggtaat ctcgaggttt
     5281 aaacgtcgac gcggccgcta ctagggacag gattggtgac agaaaagccc catccttagg
     5341 cctcctcctt cctagtctcc tgatattggg tctaaccccc acctcctgtt aggcagattc
     5401 cttatctggt gacacacccc catttcctgg agccatctct ctccttgcca gaacctctaa
     5461 ggtttgctta cgatggagcc agagaggatc ctgggaggga gagcttggca gggggtggga
     5521 gggaaggggg ggatgcgtga cctgcccggt tctcagtggc caccctgcgc taccctctcc
     5581 cagaacctga gctgctctga cgcggctgtc tggtgcgttt cactgatcct ggtgctgcag
     5641 cttccttaca cttcccaaga ggagaagcag tttggaaaaa caaaatcaga ataagttggt
     5701 cctgagttct aactttggct cttcaccttt ctagtcccca atttatattg ttcctccgtg
     5761 cgtcagtttt acctgtgaga taaggccagt agccagcccc gtcctggcag ggctgtggtg
     5821 aggagggggg tgtccgtgtg gaaaactccc tttgtgagaa tggtgcgtcc taggtgttca
     5881 ccaggtcgtg gccgcctcta ctccctttct ctttctccat ccttctttcc ttaaagagtc
     5941 cccagtgcta tctgggacat attcctccgc ccagagcagg gtcccgcttc cctaaggccc
     6001 tgctctgggc ttctgggttt gagtccttgg caagcccagg agaggcgctc aggcttccct
     6061 gtcccccttc ctcgtccacc atctcatgcc cctggctctc ctgccccttc cctacagggg
     6121 ttcctggctc tgctctccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     6181 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6241 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6301 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6361 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6421 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6481 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6541 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6601 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6661 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6721 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6781 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6841 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6901 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6961 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7021 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7081 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7141 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7201 tcctttgat
//
