LOCUS       40924_28047        7350 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo622, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7350)
  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 7350)
  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 7350)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7350)
  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..7350
                     /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..1073
                     /label=8g8f5
                     /note="8g8f5"
     misc_feature    315..647
                     /label=light
                     /note="light"
     misc_feature    708..1073
                     /label=heavy
                     /note="heavy"
     misc_feature    1086..1829
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    1359..1361
                     /label=F93A
                     /note="F93A"
     misc_feature    1839..2667
                     /label=IL6ST
                     /note="IL6ST"
     misc_feature    2190..2192
                     /label=Y759A
                     /note="Y759A"
     polyA_signal    2723..2947
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2993..3421
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3435..3764
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3831..4622
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4799..4932
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4969..4985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4993..5009)
                     /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(5017..5047)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5062..5083)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5371..5956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6130..6987)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6988..7092)
                     /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 atccgacatt gtcctgaccc aaagcccagc gtcgttagcg gtgtccctgg
      361 ggcaacgtgc tactatttcc tgcaaagcct cccaatctgt tgatttcgat ggggactcat
      421 acatgaactg gtatcagcaa aagcctggac agccgcctaa attacttatt ttcgccgcta
      481 gtaatctggc tagtggcatt cctgctcgcc tgtcaggttc aggttcgggc acggatttca
      541 cgctgaatat ccagcccgtt gaggaagaag atgcagccac atactactgc caacagtcca
      601 atgaagatcc ctacactttc ggggggggca ctaagttgga gatcaagggt ggtggaggat
      661 caggtggtgg agggtctggc ggaggcggta gcggtggagg tggaagccaa gtgcagcttc
      721 agcagagcgg agatgattta gtcaagccag gcgcaagtgt taaattatcc tgtaaagcat
      781 caggttacac attcactact tattacatca actggatgcg ccagcgccca ggacaaggat
      841 tagagtggat cggtcgtatt gctccagctt caggcacaac ttactcctca gagatgttca
      901 aggataaggc aacgttgact gttgatacat cttcgaatac agcgtacatt cagcttagct
      961 cgttgagttc tgaagatagt gccgtctatt tctgtgctcg cgctgattac ggctttaata
     1021 gcggggaggc tatggattat tggggacaag gtacttctgt cacggtgagt tcgtccggag
     1081 aattcgcacc ttcacccagc ctcccggacc ccaagtttga gagcaaagcg gccctgctgg
     1141 catcccgggg ctccgaagaa cttctgtgct tcacccaacg cttggaagac ttggtgtgtt
     1201 tctgggagga agcggcgagc tccgggatgg acttcaacta cagcttctca taccagctcg
     1261 agggtgagtc acgaaagtca tgtagcctgc accaggctcc caccgtccgc ggctccgtgc
     1321 gtttctggtg ttcactgcca acagcggaca catcgagtgc tgtgccgctg gagctgcagg
     1381 tgacggaggc gtccggttct cctcgctatc accgcatcat ccatatcaat gaagtagtgc
     1441 tcctggacgc ccccgcgggg ctgctggcgc gccgggcaga agagggcagc cacgtggtgc
     1501 tgcgctggct gccacctcct ggagcaccta tgaccaccca catccgatat gaagtggacg
     1561 tgtcggcagg caaccgggca ggagggacac aaagggtgga ggtcctggaa ggccgcactg
     1621 agtgtgttct gagcaacctg cggggcggga cgcgctacac cttcgctgtt cgagcgcgca
     1681 tggccgagcc gagcttcagc ggattctgga gtgcctggtc tgagcccgcg tcactactga
     1741 ccgctagcga cctggaccct ctcatcttga cgctgtctct cattctggtc ctcatctcgt
     1801 tgttgctgac ggttctggcc ctgctgtccg cagcagcaaa taagcgagac ctaattaaaa
     1861 aacacatctg gcctaatgtt ccagatcctt caaagagtca tattgcccag tggtcacctc
     1921 acactcctcc aaggcacaat tttaattcaa aagatcaaat gtattcagat ggcaatttca
     1981 ctgatgtaag tgttgtggaa atagaagcaa atgacaaaaa gccttttcca gaagatctga
     2041 aatcattgga cctgttcaaa aaggaaaaaa ttaatactga aggacacagc agtggtattg
     2101 gggggtcttc atgcatgtca tcttctaggc caagcatttc tagcagtgat gaaaatgaat
     2161 cttcacaaaa cacttcgagc actgtccagg cttctaccgt ggtacacagt ggctacagac
     2221 accaagttcc gtcagtccaa gtcttctcaa gatccgagtc tacccagccc ttgttagatt
     2281 cagaggagcg gccagaagat ctacaattag tagatcatgt agatggcggt gatggtattt
     2341 tgcccaggca acagtacttc aaacagaact gcagtcagca tgaatccagt ccagatattt
     2401 cacattttga aaggtcaaag caagtttcat cagtcaatga ggaagatttt gttagactta
     2461 aacagcagat ttcagatcat atttcacaat cctgtggatc tgggcaaatg aaaatgtttc
     2521 aggaagtttc tgcagcagat gcttttggtc caggtactga gggacaagta gaaagatttg
     2581 aaacagttgg catggaggct gcgactgatg aaggcatgcc taaaagttac ttaccacaga
     2641 ctgtacggca aggcggctac atgcctcagt aggcggccgc tcgagtctag agggcccgtt
     2701 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2761 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2821 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2881 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2941 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3001 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3061 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3121 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3181 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3241 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3301 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3361 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3421 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3481 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3541 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3601 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3661 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3721 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3781 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3841 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3901 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3961 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4021 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4081 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4141 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4201 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4261 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4321 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4381 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4441 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4501 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4561 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4621 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4681 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4741 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4801 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4861 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4921 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4981 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5041 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5101 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5161 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5221 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5281 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5341 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5401 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5461 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5521 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5581 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5641 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5701 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5761 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5821 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5881 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     5941 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6001 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6061 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6121 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6181 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6241 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6301 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6361 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6421 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6481 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6541 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6601 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6661 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6721 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6781 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6841 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6901 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6961 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7021 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7081 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7141 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     7201 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7261 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     7321 gcgctgcttc gcgatgtacg ggccagatat
//
