LOCUS       40924_18841        8784 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pEXPR1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8784)
  AUTHORS   Siciliano V.
  TITLE     Engineering modular intracellular protein sensor-actuator devices
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8784)
  AUTHORS   Siciliano V.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-APR-2018) Biological Engineering, Massachusetts 
            Institute of Technology, 500 Tech Square, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 8784)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8784)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-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..8784
                     /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"
     promoter        4252..5424
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     protein_bind    5446..5470
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5476..6165
                     /codon_start=1
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGH"
     protein_bind    complement(6817..6841)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6996..7051
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7412..7428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7436..7452
                     /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(7460..7490)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7505..7526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    7699..8535
                     /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(8565..8583)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8604..8620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8628..8644)
                     /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(8652..8682)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8697..8718)
                     /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 ccgaattcgc ccttattaat tggctccggt
     4261 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttgtg gggaggggtc
     4321 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     4381 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     4441 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
     4501 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
     4561 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
     4621 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
     4681 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
     4741 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
     4801 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
     4861 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     4921 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     4981 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     5041 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     5101 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     5161 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     5221 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     5281 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     5341 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     5401 agtttttttc ttccatttca ggtggatccc aagggcgaat tcgacccaag tttgtacaaa
     5461 aaagcaggct ccaccatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
     5521 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
     5581 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
     5641 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
     5701 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
     5761 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     5821 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
     5881 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     5941 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag
     6001 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     6061 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     6121 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacggcgg aggtggatct
     6181 ggcggaggtg gatcgggcgg aggtggatca atggcgccta tcggctcagt agtaatcgta
     6241 ggcagaatca tcctgtccgg ccgtggtggc ccgatcacgg cctactccca acagacgcgg
     6301 ggcctacttg gttgcattat cactagtctc acgggccggg acaagaacca ggttgagggg
     6361 gaggttcaag tggtctccac cgcaacacaa tctttcctgg cgacctgtgt caacggcgcg
     6421 tgctggactg tctttcacgg tgccggctca aagaccttag ccggcccaaa aggcccaatc
     6481 acccaaatgt acaccaatgt agaccaggac ctcgtcggct ggccagcgcc ccccggggcg
     6541 cgttccttaa caccgtgcac ctgcggcagc tcggaccttt acctggtcac gagacatgct
     6601 gatgtcatcc cggtgcgccg gcggggcgac actaggggga gcttgctctc ccctagaccc
     6661 atctcctact tgaagggctc ttcgggtggt ccattgctct gcccctcggg gcacgttgtg
     6721 ggcatcttcc gggctgccgt gtgcacccgg ggggtcgcga aggcggtgga cttcataccc
     6781 gttgaatcta tggaaaccac tatgcggggc tcctaaaccc agctttcttg tacaaagtgg
     6841 tacgcgtgaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     6901 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     6961 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     7021 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     7081 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     7141 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     7201 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     7261 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     7321 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     7381 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7441 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7501 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7561 gaaacctgtc gtgccagcgg atcgacagta ctaagcttgg tgcgttttta tgcttgtagt
     7621 attgtataat gtttttaaga tccttaatta atagggataa cagggtaatc tcgaggttta
     7681 aacgtcgacg cggccgctac tagggacagg attggtgaca gaaaagcccc atccttaggc
     7741 ctcctccttc ctagtctcct gatattgggt ctaaccccca cctcctgtta ggcagattcc
     7801 ttatctggtg acacaccccc atttcctgga gccatctctc tccttgccag aacctctaag
     7861 gtttgcttac gatggagcca gagaggatcc tgggagggag agcttggcag ggggtgggag
     7921 ggaagggggg gatgcgtgac ctgcccggtt ctcagtggcc accctgcgct accctctccc
     7981 agaacctgag ctgctctgac gcggctgtct ggtgcgtttc actgatcctg gtgctgcagc
     8041 ttccttacac ttcccaagag gagaagcagt ttggaaaaac aaaatcagaa taagttggtc
     8101 ctgagttcta actttggctc ttcacctttc tagtccccaa tttatattgt tcctccgtgc
     8161 gtcagtttta cctgtgagat aaggccagta gccagccccg tcctggcagg gctgtggtga
     8221 ggaggggggt gtccgtgtgg aaaactccct ttgtgagaat ggtgcgtcct aggtgttcac
     8281 caggtcgtgg ccgcctctac tccctttctc tttctccatc cttctttcct taaagagtcc
     8341 ccagtgctat ctgggacata ttcctccgcc cagagcaggg tcccgcttcc ctaaggccct
     8401 gctctgggct tctgggtttg agtccttggc aagcccagga gaggcgctca ggcttccctg
     8461 tcccccttcc tcgtccacca tctcatgccc ctggctctcc tgccccttcc ctacaggggt
     8521 tcctggctct gctctccacc gcggtggagc tccagctttt gttcccttta gtgagggtta
     8581 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     8641 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     8701 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     8761 tcgtgccagc tgcattaatg aatc
//
