LOCUS       40924_18876        7836 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pEXPR38, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7836)
  AUTHORS   Siciliano V.
  TITLE     Engineering modular intracellular protein sensor-actuator devices
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7836)
  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 7836)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7836)
  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            "Unpublished"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..7836
                     /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"
     protein_bind    complement(5869..5893)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6048..6103
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6464..6480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6488..6504
                     /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(6512..6542)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6557..6578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    6751..7587
                     /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(7617..7635)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7656..7672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7680..7696)
                     /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(7704..7734)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7749..7770)
                     /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 gccagggcgg aggtggatct ggcggaggtg gatcgggcgg aggtggatca
     5461 atggccgagg tccaggtgca gcagtctggg gcagagcttg tgaggtcagg ggcctcagtc
     5521 aagttgtcct gcacagcttc tggcttcaac attaaagact actatatgca ctgggtgaag
     5581 cagaggcctg aacagggcct ggagtggatt ggatggattg atcctgcgaa tggtaatact
     5641 aaatatgacc cgaagttcca gggcaaggcc actataacag cagacacatc ctccaacaca
     5701 gcctacctgc agctcagcag cctgacatct gaggacactg ccgtctatta ctgtgctaga
     5761 tgggactggt acttcgatgt ctggggcgca gggaccacag tcacagtctc ctcgggaggt
     5821 ggtggatccg gcggtggcgg ttctggtgga ggtggatctg tgttttgaac ccagctttct
     5881 tgtacaaagt ggtacgcgtg aattcactcc tcaggtgcag gctgcctatc agaaggtggt
     5941 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     6001 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     6061 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     6121 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     6181 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     6241 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     6301 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     6361 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     6421 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     6481 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6541 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6601 ctttccagtc gggaaacctg tcgtgccagc ggatcgacag tactaagctt ggtctaatac
     6661 ccaatctctc gtcttatcca gatgttttat acgccttaat taatagggat aacagggtaa
     6721 tctcgaggtt taaacgtcga cgcggccgct actagggaca ggattggtga cagaaaagcc
     6781 ccatccttag gcctcctcct tcctagtctc ctgatattgg gtctaacccc cacctcctgt
     6841 taggcagatt ccttatctgg tgacacaccc ccatttcctg gagccatctc tctccttgcc
     6901 agaacctcta aggtttgctt acgatggagc cagagaggat cctgggaggg agagcttggc
     6961 agggggtggg agggaagggg gggatgcgtg acctgcccgg ttctcagtgg ccaccctgcg
     7021 ctaccctctc ccagaacctg agctgctctg acgcggctgt ctggtgcgtt tcactgatcc
     7081 tggtgctgca gcttccttac acttcccaag aggagaagca gtttggaaaa acaaaatcag
     7141 aataagttgg tcctgagttc taactttggc tcttcacctt tctagtcccc aatttatatt
     7201 gttcctccgt gcgtcagttt tacctgtgag ataaggccag tagccagccc cgtcctggca
     7261 gggctgtggt gaggaggggg gtgtccgtgt ggaaaactcc ctttgtgaga atggtgcgtc
     7321 ctaggtgttc accaggtcgt ggccgcctct actccctttc tctttctcca tccttctttc
     7381 cttaaagagt ccccagtgct atctgggaca tattcctccg cccagagcag ggtcccgctt
     7441 ccctaaggcc ctgctctggg cttctgggtt tgagtccttg gcaagcccag gagaggcgct
     7501 caggcttccc tgtccccctt cctcgtccac catctcatgc ccctggctct cctgcccctt
     7561 ccctacaggg gttcctggct ctgctctcca ccgcggtgga gctccagctt ttgttccctt
     7621 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     7681 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     7741 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7801 tcgggaaacc tgtcgtgcca gctgcattaa tgaatc
//
