LOCUS       sequence-211288-        6585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E.coli entry and expression vector for FX cloning system, N-terminal
            pelB signal sequence and C-terminal fusion to PIII for phage display
            using M13 phages.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6585)
  AUTHORS   Zimmermann I, Egloff P, Hutter CA, Arnold FM, Stohler P, Bocquet N, 
            Hug MN, Huber S, Siegrist M, Hetemann L, Gera J, Gmur S, Spies P, 
            Gygax D, Geertsma ER, Dawson RJ, Seeger MA
  TITLE     Synthetic single domain antibodies for the conformational trapping 
            of membrane proteins.
  JOURNAL   Elife. 2018 May 24;7. pii: 34317. doi: 10.7554/eLife.34317.
  PUBMED    29792401
REFERENCE   2  (bases 1 to 6585)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2018-05-24"; volume: "7. pii: 34317. doi"; pages: " 
            10.7554/eLife.34317"
COMMENT     Sequence Label: pDXinit
FEATURES             Location/Qualifiers
     source          1..6585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     21..86
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     promoter        107..552
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(553..639,640..1974)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     terminator      2048..2134
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             2208..2225
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3510..3527)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3510..3526)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3519..3541)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3739..4194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3826..3845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4036..4057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4211..4230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4330..4352
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4390..4408)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4476..4580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4581..4649,4650..5441)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4799..4818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5612..6200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6101..6120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6354..6371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6488..6509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6524..6529,6530..6547,6548..6554)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6562..6578
                     /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)."
ORIGIN
        1 agcttaagga gacagtacat atgaaatacc tattgcctac ggcagccgct ggattgttat
       61 tactcgcggc ccagccggcc atggcagccg gctcttcaag tctgcccaca tatacctgcc
      121 gttcactatt atttagtgaa atgagatatt atgatatttt ctgaattgtg attaaaaagg
      181 caactttatg cccatgcaac agaaactata aaaaatacag agaatgaaaa gaaacagata
      241 gattttttag ttctttaggc ccgtagtctg caaatccttt tatgattttc tatcaaacaa
      301 aagaggaaaa tagaccagtt gcaatccaaa cgagagtcta atagaatgag gtcgaaaagt
      361 aaatcgcgcg ggtttgttac tgataaagca ggcaagacct aaaatgtgta aagggcaaag
      421 tgtatacttt ggcgtcaccc cttacatatt ttaggtcttt ttttattgtg cgtaactaac
      481 ttgccatctt caaacaggag ggctggaaga agcagaccgc taacacagta cataaaaaag
      541 gagacatgaa cgatgaacat caaaaagttt gcaaaacaag caacagtatt aacctttact
      601 accgcactgc tggcaggagg cgcaactcaa gcgtttgcga aagaaacgaa ccaaaagcca
      661 tataaggaaa catacggcat ttcccatatt acacgccatg atatgctgca aatccctgaa
      721 cagcaaaaaa atgaaaaata taaagttcct gagttcgatt cgtccacaat taaaaatatc
      781 tcttctgcaa aaggcctgga cgtttgggac agctggccat tacaaaacac tgacggcact
      841 gtcgcaaact atcacggcta ccacatcgtc tttgcattag ccggagatcc taaaaatgcg
      901 gatgacacat cgatttacat gttctatcaa aaagtcggcg aaacttctat tgacagctgg
      961 aaaaacgctg gccgcgtctt taaagacagc gacaaattcg atgcaaatga ttctatccta
     1021 aaagaccaaa cacaagaatg gtcaggttca gccacattta catctgacgg aaaaatccgt
     1081 ttattctaca ctgatttctc cggtaaacat tacggcaaac aaacactgac aactgcacaa
     1141 gttaacgtat cagcatcaga cagctctttg aacatcaacg gtgtagagga ttataaatca
     1201 atctttgacg gtgacggaaa aacgtatcaa aatgtacagc agttcatcga tgaaggcaac
     1261 tacagctcag gcgacaacca tacgctgaga gatcctcact acgtagaaga taaaggccac
     1321 aaatacttag tatttgaagc aaacactgga actgaagatg gctaccaagg cgaagaatct
     1381 ttatttaaca aagcatacta tggcaaaagc acatcattct tccgtcaaga aagtcaaaaa
     1441 cttctgcaaa gcgataaaaa acgcacggct gagttagcaa acggcgctct cggtatgatt
     1501 gagctaaacg atgattacac actgaaaaaa gtgatgaaac cgctgattgc atctaacaca
     1561 gtaacagatg aaattgaacg cgcgaacgtc tttaaaatga acggcaaatg gtacctgttc
     1621 actgactccc gcggatcaaa aatgacgatt gacggcatta cgtctaacga tatttacatg
     1681 cttggttatg tttctaattc tttaactggc ccatacaagc cgctgaacaa aactggcctt
     1741 gtgttaaaaa tggatcttga tcctaacgat gtaaccttta cttactcaca cttcgctgta
     1801 cctcaagcga aaggaaacaa tgtcgtgatt acaagctata tgacaaacag aggattctac
     1861 gcagacaaac aatcaacgtt tgcgcctagc ttcctgctga acatcaaagg caagaaaaca
     1921 tctgttgtca aagacagcat ccttgaacaa ggacaattaa cagttaacaa ataaaaacgc
     1981 aaaagaaaat gccgatatcc tattggcatt gcggccgcgg tctcggcggt gggtcgtcta
     2041 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2101 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacct agcggccggc
     2161 cgcaagcgct gcacggcctc acgtgacacg tgtctgcagg aagagctcac caccatcacc
     2221 atcacgaacc tgaagcccag tacccgtacg acgttccgga ctacggttcc actgttgaaa
     2281 gttgtttagc aaaacctcat acagaaaatt catttactaa cgtctggaaa gacgacaaaa
     2341 ctttagatcg ttacgctaac tatgagggct gtctgtggaa tgctacaggc gttgtcgttt
     2401 gtactggtga cgaaactcag tgttacggta catgggttcc tattgggctt gctatccctg
     2461 aaaatgaggg tggtggctct gagggtggcg gttctgaggg tggcggttct gagggtggcg
     2521 gtactaaacc tcctgagtac ggtgatacac ctattccggg ctatacttat atcaaccctc
     2581 tcgacggcac ttatccgcct ggtactgagc aaaaccccgc taatcctaat ccttctcttg
     2641 aggagtctca gcctcttaat actttcatgt ttcagaataa taggttccga aataggcagg
     2701 gtgcattaac tgtttatacg ggcactgtta ctcaaggcac tgaccccgtt aaaacttatt
     2761 accagtacac tcctgtatca tcaaaagcca tgtatgacgc ttactggaac ggtaaattca
     2821 gagactgcgc tttccattct ggctttaatg aggatccatt cgtttgtgaa tatcaaggcc
     2881 aatcgtctga cctgcctcaa cctcctgtca atgctggcgg cggctctggt ggtggttctg
     2941 gtggcggctc tgagggtggc ggctctgagg gtggcggctc tgagggtggc ggttctgagg
     3001 gtggcggctc tgagggtggc ggttccggtg gcggctccgg ttccggtgat tttgattatg
     3061 aaaaaatggc aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac
     3121 agtctgacgc taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg
     3181 gtttcattgg tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg
     3241 gctctaattc ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt
     3301 tccgtcaata tttaccttct ttacctcagt cggttgaatg tcgcccttat gtctttggcg
     3361 ctggtaaacc atatgaattt tctattgatt gtgacaaaat aaacttattc cgtggtgtct
     3421 ttgcgtttct tttatatgtt gccaccttta tgtatgtatt ttcgacgttt gctaacatac
     3481 tgcgtaataa ggagtcttaa taagaattca ctggccgtcg ttttacaacg tcgtgactgg
     3541 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3601 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3661 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3721 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3781 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     3841 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3901 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     3961 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4021 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     4081 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4141 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4261 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccacttac agacaagctg
     4321 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4381 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4441 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4501 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4561 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4621 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4681 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4741 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4801 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4861 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4921 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4981 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5041 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5101 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5161 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5221 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5281 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5341 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5401 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5461 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5521 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5581 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5641 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5701 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5761 ttctagtgta gccgtagtta ggccactact tcaagaactc tgtagcaccg cctacatacc
     5821 tcgcactgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5881 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5941 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6001 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6061 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6181 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6241 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6301 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6361 cagtgagcga ggaagcggaa gagagcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6421 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6481 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6541 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcaca
//
