LOCUS       40924_28057        6939 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo626, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  AUTHORS   Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
  TITLE     Generalized extracellular molecule sensor platform for programming 
            cellular behavior
  JOURNAL   Nat. Chem. Biol. 14 (7), 723-729 (2018)
  PUBMED    29686358
REFERENCE   2  (bases 1 to 6939)
  AUTHORS   Scheller L, Strittmatter T.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 6939)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6939)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol."; date: "2018"; volume: "14"; issue: "7"; pages: "723-729"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058, 
            Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..215
                     /label=Psv40
                     /note="Psv40"
     promoter        18..214
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      65..200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     sig_peptide     246..308
                     /label=Ig-kappa leader
                     /note="leader sequence from mouse immunoglobulin kappa 
                     light
                     chain"
     misc_feature    315..662
                     /label=Nic_VH
                     /note="Nic_VH"
     misc_feature    675..1418
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    948..950
                     /label=F93A
                     /note="F93A"
     misc_feature    1350..1418
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1428..2256
                     /label=IL6ST
                     /note="IL6ST"
     misc_feature    1779..1781
                     /label=Y759A
                     /note="Y759A"
     polyA_signal    2312..2536
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2582..3010
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3024..3353
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3420..4211
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4388..4521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4558..4574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4582..4598)
                     /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(4606..4636)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4651..4672)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4960..5545)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5719..6576)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6577..6681)
                     /label=AmpR promoter
ORIGIN
        1 acgcgtgatc tgcgatctgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
       61 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatc gctgactaat
      121 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
      181 aggaggcttt tttggaggcc taggcttttg caaaaagctt cgaatcgcgc tagcggccgg
      241 ccaccatgga aactgatact ttgctgctct gggtcctgct gctgtgggtc cctggatcaa
      301 cgggggacgg atcccagatg caactgttgg aatcgggacc gggtttggtc aagccgagcg
      361 agacattgag tttaacgtgc acggtttcag gcggatcaat ttggggttgg attcgccagc
      421 cgccaggaaa gggactggag tggattggat ctatttactc ctccggttcc acctactata
      481 atccgagctt gaagtcccgt gtgacaacaa gcgtggatac atccaaaaac cagttttcat
      541 tgcgcctgtc ctcagtaacc gcagccgaca cagccgtata ttattgcgtt gcttggtttg
      601 gcgacttatt aagtcttaaa ggggtagagc tttggggcca gggaactctt gtgacggtat
      661 cgtccggaga attcgcacct tcacccagcc tcccggaccc caagtttgag agcaaagcgg
      721 ccctgctggc atcccggggc tccgaagaac ttctgtgctt cacccaacgc ttggaagact
      781 tggtgtgttt ctgggaggaa gcggcgagct ccgggatgga cttcaactac agcttctcat
      841 accagctcga gggtgagtca cgaaagtcat gtagcctgca ccaggctccc accgtccgcg
      901 gctccgtgcg tttctggtgt tcactgccaa cagcggacac atcgagtgct gtgccgctgg
      961 agctgcaggt gacggaggcg tccggttctc ctcgctatca ccgcatcatc catatcaatg
     1021 aagtagtgct cctggacgcc cccgcggggc tgctggcgcg ccgggcagaa gagggcagcc
     1081 acgtggtgct gcgctggctg ccacctcctg gagcacctat gaccacccac atccgatatg
     1141 aagtggacgt gtcggcaggc aaccgggcag gagggacaca aagggtggag gtcctggaag
     1201 gccgcactga gtgtgttctg agcaacctgc ggggcgggac gcgctacacc ttcgctgttc
     1261 gagcgcgcat ggccgagccg agcttcagcg gattctggag tgcctggtct gagcccgcgt
     1321 cactactgac cgctagcgac ctggaccctc tcatcttgac gctgtctctc attctggtcc
     1381 tcatctcgtt gttgctgacg gttctggccc tgctgtccgc agcagcaaat aagcgagacc
     1441 taattaaaaa acacatctgg cctaatgttc cagatccttc aaagagtcat attgcccagt
     1501 ggtcacctca cactcctcca aggcacaatt ttaattcaaa agatcaaatg tattcagatg
     1561 gcaatttcac tgatgtaagt gttgtggaaa tagaagcaaa tgacaaaaag ccttttccag
     1621 aagatctgaa atcattggac ctgttcaaaa aggaaaaaat taatactgaa ggacacagca
     1681 gtggtattgg ggggtcttca tgcatgtcat cttctaggcc aagcatttct agcagtgatg
     1741 aaaatgaatc ttcacaaaac acttcgagca ctgtccaggc ttctaccgtg gtacacagtg
     1801 gctacagaca ccaagttccg tcagtccaag tcttctcaag atccgagtct acccagccct
     1861 tgttagattc agaggagcgg ccagaagatc tacaattagt agatcatgta gatggcggtg
     1921 atggtatttt gcccaggcaa cagtacttca aacagaactg cagtcagcat gaatccagtc
     1981 cagatatttc acattttgaa aggtcaaagc aagtttcatc agtcaatgag gaagattttg
     2041 ttagacttaa acagcagatt tcagatcata tttcacaatc ctgtggatct gggcaaatga
     2101 aaatgtttca ggaagtttct gcagcagatg cttttggtcc aggtactgag ggacaagtag
     2161 aaagatttga aacagttggc atggaggctg cgactgatga aggcatgcct aaaagttact
     2221 taccacagac tgtacggcaa ggcggctaca tgcctcagta ggcggccgct cgagtctaga
     2281 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2341 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2401 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2461 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2521 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2581 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2641 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2701 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2761 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2821 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2881 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2941 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3001 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3061 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3181 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3241 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3301 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3361 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3421 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3481 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3541 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3601 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3661 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3721 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3781 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3841 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3901 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3961 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4021 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4081 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4141 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4201 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4261 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4321 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4381 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4441 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4501 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4561 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4621 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4681 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4741 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4801 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4861 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4921 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4981 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5041 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5101 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5161 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5221 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5281 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5341 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5401 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5461 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     5521 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5581 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5641 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5701 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5761 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5821 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5881 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5941 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6001 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6061 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6121 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6181 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6241 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6301 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6361 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6421 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6481 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6541 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6601 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6661 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6721 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6781 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6841 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     6901 aggcgttttg cgctgcttcg cgatgtacgg gccagatat
//
