LOCUS       sequence-177298-        5985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express csgB curli protein with GS linker and Histaq.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5985)
  AUTHORS   Abdelwahab MT, Kalyoncu E, Onur T, Baykara MZ, Seker UO
  TITLE     Genetically-Tunable Mechanical Properties of Bacterial Functional 
            Amyloid Nanofibers.
  JOURNAL   Langmuir. 2017 Apr 7. doi: 10.1021/acs.langmuir.7b00112.
  PUBMED    28388843
REFERENCE   2  (bases 1 to 5985)
  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: "Langmuir. 
            2017 Apr 7. doi: 10.1021/acs.langmuir.7b00112."
COMMENT     Sequence Label: pET22b-csgB
FEATURES             Location/Qualifiers
     source          1..5985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(60..851,852..920))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     683..702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(921..1025)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1051..1506)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1188..1209)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1400..1419
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(1543..1590)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1586..1604
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1657..1674)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1717..1737)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     sig_peptide     complement(2237..2302)
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     RBS             2310..2332
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2347..2371
                     /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)."
     primer_bind     complement(2371..2390)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2372..2390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2474..2493)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2699..2776
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2777..3859
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2796..2815)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4668..4859
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4853..4875)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4961..5100
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5016..5035
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5115..5132)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5286..5874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5366..5385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
       61 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
      121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
      181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
      241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
      301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
      361 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
      421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
      481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
      541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
      601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
      661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
      721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
      781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
      841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
      901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
      961 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     1021 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     1081 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1141 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1201 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1261 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1321 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1381 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1441 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1501 gcgcgtccca ttcgccaatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac
     1561 ccgtttagag gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc
     1621 agcttccttt cgggctttgt tagcagccgg atctcagtgg tggtggtggt ggtgctcgag
     1681 gccgctaccg cctccgctac cgccaccgct accgccgccc tgaaaataca ggttttcgcc
     1741 gctaccgcct ccgctaccgc caccgctacc gccggtacca cgttgtgtca cgcgaatagc
     1801 catttgcgac tgtctctgca ctacaattgc cgttttttga gtaccatact gtgtaatatt
     1861 tgctttatta ccagaacctt tctggataat catcgcagta ttaccataag caccttgcga
     1921 aatactggca tcgttggcac tgcccgcctg atcaatatat gcaaggttat aatctcctgt
     1981 ctggtcaatc tttgcccggt tgctactacc ttcttgcgca acaaccgcca aaagttttga
     2041 gcctccctgc cgtaactgag cactattatt agtcccagct tgaccaatta tggctgcctg
     2101 attaaatgaa gacttactca attcatttac cgcgaagtta tattctgaat tagctaaatc
     2161 ataacctgct gcggctgcaa tcccaggcgc acccagtatt gttaacatca taaataacaa
     2221 tttgtttttc atccatggcc atcgccggct gggcagcgag gagcagcaga ccagcagcag
     2281 cggtcggcag caggtatttc atatgtatat ctccttctta aagttaaaca aaattatttc
     2341 tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg
     2401 atcgagatct cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     2461 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     2521 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     2581 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
     2641 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga
     2701 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     2761 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     2821 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     2881 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     2941 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     3001 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     3061 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     3121 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     3181 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     3241 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     3301 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     3361 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     3421 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     3481 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     3541 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
     3601 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     3661 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     3721 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     3781 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3841 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     3901 cgggatctcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     3961 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     4021 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     4081 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     4141 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc
     4201 cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc
     4261 ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca
     4321 aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc
     4381 tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct
     4441 gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag
     4501 tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca
     4561 gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg
     4621 gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg
     4681 aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga
     4741 aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg
     4801 ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     4861 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     4921 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     4981 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     5041 agtgcaccat tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     5101 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5161 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5221 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5281 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5341 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5401 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5461 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5521 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5581 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5641 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5701 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5761 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5821 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5881 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5941 ttggtcatga gattatcaaa aaggatcttc acctagatcc tttta
//
