LOCUS       40924_28037        7323 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo620, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7323)
  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 7323)
  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 7323)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7323)
  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..7323
                     /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    309..1043
                     /label=scfv
                     /note="scfv"
     misc_feature    1059..1802
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    1332..1334
                     /label=F93A
                     /note="F93A"
     misc_feature    1812..2640
                     /label=IL6ST
                     /note="IL6ST"
     misc_feature    2163..2165
                     /label=Y759A
                     /note="Y759A"
     polyA_signal    2696..2920
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2966..3394
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3408..3737
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3804..4595
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4772..4905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4942..4958)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4966..4982)
                     /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(4990..5020)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5035..5056)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5344..5929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6103..6960)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6961..7065)
                     /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 atccgacatc gtgatgaccc agagccccag cagcctgagc gccagcgtgg
      361 gcgaccgcgt gaccatcacc tgccgcagca gcaccggcgc cgtgaccacc agcaactacg
      421 ccagctgggt gcaggagaag cccggcaagc tgttcaaggg cctgatcggc ggcaccaaca
      481 accgcgcccc cggcgtgccc agccgcttca gcggcagcct gatcggcgac aaggccaccc
      541 tgaccatcag cagcctgcag cccgaggact tcgccaccta cttctgcgcc ctgtggtaca
      601 gcaaccactg ggtgttcggc cagggcacca aggtggagct gaagcgcggc ggcggcggca
      661 gcggcggcgg cggcagcggc ggcggcggca gcagcggcgg cggcagcgag gtgaagctgc
      721 tggagagcgg cggcggcctg gtgcagcccg gcggcagcct gaagctgagc tgcgccgtga
      781 gcggcttcag cctgaccgac tacggcgtga actgggtgcg ccaggccccc ggccgcggcc
      841 tggagtggat cggcgtgatc tggggcgacg gcatcaccga ctacaacagc gccctgaagg
      901 accgcttcat catcagcaag gacaacggca agaacaccgt gtacctgcag atgagcaagg
      961 tgcgcagcga cgacaccgcc ctgtactact gcgtgaccgg cctgttcgac tactggggcc
     1021 agggcaccct ggtgaccgtg agcagctccg gagaattcgc accttcaccc agcctcccgg
     1081 accccaagtt tgagagcaaa gcggccctgc tggcatcccg gggctccgaa gaacttctgt
     1141 gcttcaccca acgcttggaa gacttggtgt gtttctggga ggaagcggcg agctccggga
     1201 tggacttcaa ctacagcttc tcataccagc tcgagggtga gtcacgaaag tcatgtagcc
     1261 tgcaccaggc tcccaccgtc cgcggctccg tgcgtttctg gtgttcactg ccaacagcgg
     1321 acacatcgag tgctgtgccg ctggagctgc aggtgacgga ggcgtccggt tctcctcgct
     1381 atcaccgcat catccatatc aatgaagtag tgctcctgga cgcccccgcg gggctgctgg
     1441 cgcgccgggc agaagagggc agccacgtgg tgctgcgctg gctgccacct cctggagcac
     1501 ctatgaccac ccacatccga tatgaagtgg acgtgtcggc aggcaaccgg gcaggaggga
     1561 cacaaagggt ggaggtcctg gaaggccgca ctgagtgtgt tctgagcaac ctgcggggcg
     1621 ggacgcgcta caccttcgct gttcgagcgc gcatggccga gccgagcttc agcggattct
     1681 ggagtgcctg gtctgagccc gcgtcactac tgaccgctag cgacctggac cctctcatct
     1741 tgacgctgtc tctcattctg gtcctcatct cgttgttgct gacggttctg gccctgctgt
     1801 ccgcagcagc aaataagcga gacctaatta aaaaacacat ctggcctaat gttccagatc
     1861 cttcaaagag tcatattgcc cagtggtcac ctcacactcc tccaaggcac aattttaatt
     1921 caaaagatca aatgtattca gatggcaatt tcactgatgt aagtgttgtg gaaatagaag
     1981 caaatgacaa aaagcctttt ccagaagatc tgaaatcatt ggacctgttc aaaaaggaaa
     2041 aaattaatac tgaaggacac agcagtggta ttggggggtc ttcatgcatg tcatcttcta
     2101 ggccaagcat ttctagcagt gatgaaaatg aatcttcaca aaacacttcg agcactgtcc
     2161 aggcttctac cgtggtacac agtggctaca gacaccaagt tccgtcagtc caagtcttct
     2221 caagatccga gtctacccag cccttgttag attcagagga gcggccagaa gatctacaat
     2281 tagtagatca tgtagatggc ggtgatggta ttttgcccag gcaacagtac ttcaaacaga
     2341 actgcagtca gcatgaatcc agtccagata tttcacattt tgaaaggtca aagcaagttt
     2401 catcagtcaa tgaggaagat tttgttagac ttaaacagca gatttcagat catatttcac
     2461 aatcctgtgg atctgggcaa atgaaaatgt ttcaggaagt ttctgcagca gatgcttttg
     2521 gtccaggtac tgagggacaa gtagaaagat ttgaaacagt tggcatggag gctgcgactg
     2581 atgaaggcat gcctaaaagt tacttaccac agactgtacg gcaaggcggc tacatgcctc
     2641 agtaggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     2701 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2761 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2821 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2881 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2941 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3001 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3061 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3121 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3181 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3241 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3301 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3361 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3421 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3481 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3541 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3601 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3661 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3721 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3781 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3841 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3901 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3961 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4021 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4081 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4141 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4201 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4261 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4321 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4381 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4441 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4501 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4561 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4621 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4681 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4741 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4801 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4861 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4921 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4981 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5041 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5101 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5161 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5221 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5281 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5521 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5761 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5881 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5941 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6001 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6061 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6121 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6181 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6241 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6301 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6361 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6421 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6481 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6541 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6601 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6661 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6721 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6781 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6841 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6901 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6961 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7021 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7081 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     7141 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7201 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7261 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     7321 tat
//
