LOCUS       sequence-211280-        4642 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 myc and 6xHisTag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4642)
  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 4642)
  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: pSBinit
FEATURES             Location/Qualifiers
     source          1..4642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     7..69
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="KYLLPTAAAGLLLLAAQPAMA"
     CDS             complement(120..425)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(222..241)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             755..784
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             800..817
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(873..890)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     promoter        1154..1256
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1257..1916
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1323..1342)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2242..2830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2731..2750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2984..3001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3016..3156
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3082..3101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3242..3264
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3420..4298)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4325..4609
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4532..4551
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 atgagtaaat atctgctgcc gaccgcagca gcgggtctgc tgctgctggc agcccagcct
       61 gcaatggccg gctcttcaag tcgacctgca gactggctgt gtataaggga gcctgacatt
      121 tatattcccc agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga
      181 tcagccactt cttccccgat aacggagacc ggcacactgg ccatatcggt ggtcatcatg
      241 cgccagcttt catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac
      301 agcagacgtg cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca
      361 ctctgtacat ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac
      421 tgcatttcac cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc
      481 gacctcagcc atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc
      541 ttcattctgc atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc
      601 aactgatagc tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt
      661 ttgacatact tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca
      721 aatacgcata ctgttatctg caggaagagc tggcgaacaa aaactcatct cagaagagga
      781 tctgaatagc gccgtcgacc atcatcatca tcatcattga gtttaaacgg tctccagctt
      841 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
      901 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
      961 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgaa
     1021 ttccccgcgc gcgatgccct ttcgtcttcg aataaatacc tgtgacggaa gatcacttcg
     1081 cagaataaat aaatcctggt gtccctgttg ataccgggaa gccctgggcc aacttttggc
     1141 gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
     1201 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     1261 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     1321 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     1381 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     1441 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc
     1501 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     1561 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     1621 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     1681 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcga
     1741 atatggacaa cttcttcgcc cccgttttca ctatgggcaa atattatacg caaggcgaca
     1801 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     1861 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt
     1921 tttaaggcag ttattggtgc ccttaaacgc ctggttgcta cgcctgaata agtgataata
     1981 agcggatgaa tggcagaaat tcgagcccgc ctaatgagcg ggcttttttt tagcccgcct
     2041 aatgagcggg cttttttttc gaaagcaaat tcgacccatc gcgcgcgggg agtcaactca
     2101 gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatcgctg atgttacatt
     2161 gcacctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2221 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2281 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2341 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2401 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2461 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2521 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2581 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2641 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2701 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2761 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2821 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2881 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2941 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3001 ggaagcggaa gtgcccctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3061 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     3121 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     3181 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3241 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag
     3301 atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca
     3361 gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat
     3421 tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc
     3481 gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca
     3541 acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg
     3601 ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga
     3661 tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg
     3721 ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt
     3781 ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt
     3841 atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac
     3901 aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat
     3961 attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg
     4021 tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac
     4081 agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga
     4141 ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa
     4201 tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag
     4261 tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc
     4321 agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg
     4381 cacttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg
     4441 accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac
     4501 attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta
     4561 gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt
     4621 gggctaacag gaggaattaa cc
//
