LOCUS       40924_18871        7791 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pEXPR37, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7791)
  AUTHORS   Siciliano V.
  TITLE     Engineering modular intracellular protein sensor-actuator devices
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7791)
  AUTHORS   Siciliano V.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAR-2018) Biological Engineering, Massachusetts 
            Institute of Technology, 500 Tech Square, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 7791)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7791)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-MAR-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..7791
                     /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)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     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"
     protein_bind    4294..4312
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4329..4347
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4365..4383
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4401..4419
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4436..4454
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4472..4490
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        4502..4540
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    4580..4604
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4660..4684
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4696..5397
                     /codon_start=1
                     /label=TEV protease
                     /note="tobacco etch virus protease (Kapust et al., 2001)"
                     /translation="SLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKHL
                     FRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQRE
                     ERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIVG
                     IHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEEP
                     FQPVKEATQLMN"
     misc_feature    5416..5820
                     /label=Region: scFv162
                     /note="Region: scFv162"
     protein_bind    complement(5824..5848)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6003..6058
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6419..6435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6443..6459
                     /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(6467..6497)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6512..6533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    6706..7542
                     /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(7572..7590)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7611..7627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7635..7651)
                     /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(7659..7689)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7704..7725)
                     /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 tgcgttttta tgcttgtagt
     3481 attgtataat gtttttaaga tccaagcttc 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 ccgaattcgc ccttattaag ctccgaattc
     4261 gcccttcagg tccgaggttc tagacgagtt tactccctat cagtgataga gaacgatgtc
     4321 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
     4381 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttatccct
     4441 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg
     4501 aggtaggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat
     4561 cgcaaagggc gaattcgacc caagtttgta caaaaaagca ggctggatcc cggtaccctc
     4621 gacacctgct gacaggtgga tcccaagggc gaattcgacc caagtttgta caaaaaagca
     4681 ggctccgcca ccatgagcct gttcaagggc cccagagact acaaccctat cagcagcacc
     4741 atctgccacc tgaccaacga gagcgacggc cacaccacca gcctgtacgg catcggcttc
     4801 ggccccttca tcatcaccaa caagcacctg ttccggcgaa acaacggcac cctgctggtg
     4861 cagagcctgc acggcgtgtt caaagtgaag aataccacaa ccctgcagca gcacctgatc
     4921 gacggccggg acatgatcat catccggatg cccaaggact tcccaccctt cccacagaag
     4981 ctgaagttca gagagcccca gcgcgaggaa cggatctgtc tggtcacaac aaacttccag
     5041 accaagagca tgagcagcat ggtgtccgat accagctgca ccttccccag cagcgacggc
     5101 atcttctgga agcactggat tcagaccaag gacggccagt gcggcagtcc actggtgtcc
     5161 acccgggatg gcttcatcgt gggcatccac agcgccagca acttcaccaa caccaacaac
     5221 tacttcacca gcgtgcccaa gaacttcatg gaactgctga ccaaccagga agcccagcag
     5281 tgggtctccg gctggcggct gaacgccgat tctgtgctgt ggggcggaca caaggtgttc
     5341 atggtgaagc ctgaggaacc cttccagccc gtgaaagagg ccacccagct gatgaacgag
     5401 ctggtgtaca gccagatggc cgaggtccag gtgcagcagt ctggggcaga gcttgtgagg
     5461 tcaggggcct cagtcaagtt gtcctgcaca gcttctggct tcaacattaa agactactat
     5521 atgcactggg tgaagcagag gcctgaacag ggcctggagt ggattggatg gattgatcct
     5581 gcgaatggta atactaaata tgacccgaag ttccagggca aggccactat aacagcagac
     5641 acatcctcca acacagccta cctgcagctc agcagcctga catctgagga cactgccgtc
     5701 tattactgtg ctagatggga ctggtacttc gatgtctggg gcgcagggac cacagtcaca
     5761 gtctcctcgg gaggtggtgg atccggcggt ggcggttctg gtggaggtgg atctgtgttt
     5821 tgaacccagc tttcttgtac aaagtggtac gcgtgaattc actcctcagg tgcaggctgc
     5881 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     5941 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     6001 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     6061 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     6121 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
     6181 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     6241 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     6301 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     6361 tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt
     6421 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6481 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6541 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc gacagtacta
     6601 agcttggtct aatacccaat ctctcgtctt atccagatgt tttatacgcc ttaattaata
     6661 gggataacag ggtaatctcg aggtttaaac gtcgacgcgg ccgctactag ggacaggatt
     6721 ggtgacagaa aagccccatc cttaggcctc ctccttccta gtctcctgat attgggtcta
     6781 acccccacct cctgttaggc agattcctta tctggtgaca cacccccatt tcctggagcc
     6841 atctctctcc ttgccagaac ctctaaggtt tgcttacgat ggagccagag aggatcctgg
     6901 gagggagagc ttggcagggg gtgggaggga agggggggat gcgtgacctg cccggttctc
     6961 agtggccacc ctgcgctacc ctctcccaga acctgagctg ctctgacgcg gctgtctggt
     7021 gcgtttcact gatcctggtg ctgcagcttc cttacacttc ccaagaggag aagcagtttg
     7081 gaaaaacaaa atcagaataa gttggtcctg agttctaact ttggctcttc acctttctag
     7141 tccccaattt atattgttcc tccgtgcgtc agttttacct gtgagataag gccagtagcc
     7201 agccccgtcc tggcagggct gtggtgagga ggggggtgtc cgtgtggaaa actccctttg
     7261 tgagaatggt gcgtcctagg tgttcaccag gtcgtggccg cctctactcc ctttctcttt
     7321 ctccatcctt ctttccttaa agagtcccca gtgctatctg ggacatattc ctccgcccag
     7381 agcagggtcc cgcttcccta aggccctgct ctgggcttct gggtttgagt ccttggcaag
     7441 cccaggagag gcgctcaggc ttccctgtcc cccttcctcg tccaccatct catgcccctg
     7501 gctctcctgc cccttcccta caggggttcc tggctctgct ctccaccgcg gtggagctcc
     7561 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     7621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7681 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat c
//
