LOCUS       sequence-317076-        4246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pro-concanavalin A, a lectin from Canavalia ensiformis, in
            E. coli. Includes N-terminal TEV recognition site. Native N-terminal
            sequence is reconstituted upon TEV cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4246)
  AUTHORS   Nonis SG, Haywood J, Schmidberger JW, Bond CS, Mylne JS
  TITLE     Structural basis for a natural circular permutation in proteins.
  JOURNAL   bioRxiv. 2020.
REFERENCE   2  (bases 1 to 4246)
  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: "bioRxiv. 
            2020."
COMMENT     Sequence Label: pQE30-TEV-pro-conA
FEATURES             Location/Qualifiers
     source          1..4246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /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)."
     protein_bind    53..69
                     /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)."
     RBS             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             118..135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             142..162
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      984..1078
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1122..1781
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1188..1207)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1846..1932
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1979..2001)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2087..2227
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2142..2161
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2242..2259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2413..3001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2493..2512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3172..3963,3964..4032))
                     /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     3795..3814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4033..4137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4205..4223
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat taactatgag aggatcgcat
      121 caccatcacc atcacggatc cgagaatctg tatttccaaa gcacccacga gaccaatgcg
      181 ctgcacttca tgttcaacca gtttagcaag gaccaaaagg acctgattct gcaaggtgat
      241 gcgaccaccg gtaccgacgg caacctggag ctgacccgtg tgagcagcaa cggtagcccg
      301 caaggcagca gcgtgggtcg tgcgctgttc tacgcgccgg ttcacatctg ggagagcagc
      361 gcggtggttg cgagctttga agcgaccttc acctttctga ttaagagccc ggatagccac
      421 ccggcggatg gtatcgcgtt ctttattagc aacatcgaca gcagcattcc gagcggtagc
      481 accggccgtc tgctgggtct gttcccggat gcgaacgtga ttcgtaacag caccaccatc
      541 gattttaacg cggcgtataa cgcggacacc atcgtggcgg ttgaactgga tacctacccg
      601 aacaccgata ttggcgaccc gagctatccg cacatcggta ttgacatcaa gagcgttcgt
      661 agcaagaaaa ccgcgaagtg gaacatgcag aacggtaaag tgggcaccgc gcacatcatt
      721 tacaacagcg ttgacaaacg tctgagcgcg gtggttagct acccgaacgc ggatagcgcg
      781 accgtgagct atgacgttga tctggacaac gttctgccgg agtgggtgcg tgttggcctg
      841 agcgcgagca ccggtctgta taaagaaacc aacaccattc tgagctggag ctttaccagc
      901 aaactgaaga gcaatgaaat cccggacatc gcgaccgttg tgtaataagt cgacctgcag
      961 ccaagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga cctcagaact
     1021 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc
     1081 aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     1141 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     1201 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     1261 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     1321 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     1381 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     1441 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     1501 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     1561 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     1621 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     1681 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     1741 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
     1801 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
     1861 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     1921 agtaggacaa atccgccctc tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     1981 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     2041 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     2101 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     2161 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2221 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2821 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4141 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4201 taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagaaa
//
