LOCUS       sequence-387259-        5885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of SpyTag-ELP-WSCP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5885)
  TITLE     Sun lab protein expression plasmids
REFERENCE   2  (bases 1 to 5885)
  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"
COMMENT     Sequence Label: pQE80l AEW
FEATURES             Location/Qualifiers
     source          1..5885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(82..96,97..102,103..119,120..126)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    102..118
                     /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    134..150
                     /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             169..180
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             202..219
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             226..264
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     CDS             1312..1350
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     terminator      1414..1508
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1552..2211
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1618..1637)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2276..2362
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2455..3537)
                     /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     3499..3518
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3538..3615)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3699..3721)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3807..3947
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3862..3881
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3962..3979)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4133..4721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4213..4232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4892..5683,5684..5752))
                     /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     5515..5534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5753..5857)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtcttc accttgagaa atcataaaaa atttatttgc tttgtgagcg gataacaatt
      121 ataatagatt caattgtgag cggataacaa tttcacacag aattcattaa agaggagaaa
      181 ttaactatga aaggcagcag ccatcatcat catcatcacg tcgacgccca tattgtcatg
      241 gttgatgcat acaagccgac gaagctcgac ggccacggcg tgggtgttcc gggcgtcggt
      301 gtgccgggtg tgggtgtgcc gggcgaaggt gtgccgggcg tcggtgtgcc gggtgttggt
      361 gttccgggcg ttggtgtgcc gggcgttggc gtgccgggcg agggtgtgcc gggcgttggt
      421 gttccgggcg tgggtgtgcc gggcgtgggc gtgccgggcg tcggtgttcc gggcgagggt
      481 gtgccgggcg taggtgtgcc gggcgttggt gagctcatca atgatgagga acctgtcaag
      541 gacacaaacg gaaaccctct taagatagag accagatact tcatccaacc agcttcagac
      601 aataacggag gtggtcttgt cccagcaaac gttgacttat cacatttgtg tccattaggt
      661 attgtccgaa caagccttcc atatcaacca ggcctacctg ttactatctc cactccatca
      721 tcctctgaag gcaatgatgt cttgacaaac accaacatag ccatcacgtt cgatgcacca
      781 atctggcctt gtccttcttc caagacatgg acagtcgatt cctcctccga agagaagtac
      841 ataataactg gtggtgaccc taaaagtgga gaaagctttt tcaggatcga gaaatatggt
      901 aatggaaaaa acacatacaa gctcgttcgt tatgataatg gcgagggtaa gagtgttgga
      961 tccactaaga gcttgtgggg acccgctctt gttcttaatg atgatgatga tagtgatgag
     1021 aatgctttcc cgattaagtt ccgtgaggtt gacacataaa ctagtgtgcc gggcgtcggc
     1081 gtgccgggcg taggtgttcc gggcgagggt gttccgggcg ttggtgtgcc gggcgtcggc
     1141 gtgccgggcg tgggtgttcc gggcgtaggt gtgccgggcg agggtgtgcc gggcgtgggc
     1201 gtgccgggcg taggtgttcc gggcgtaggt gttccgggcg taggtgttcc gggtgaaggc
     1261 gtgccgggcg ttggtgtgcc gggtgtgggc gtgccgggcg ggctgctcga cgcccatatt
     1321 gtcatggttg atgcatacaa gccgacgaag ctcgagtgga agaaataagg atccggctgc
     1381 taacaaagcc cgaagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga
     1441 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
     1501 gtgagaatcc aagccagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
     1561 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     1621 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     1681 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1741 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     1801 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1861 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1921 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1981 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     2041 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     2101 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     2161 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     2221 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
     2281 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     2341 cgctctcctg agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc
     2401 aaacatgaga attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     2461 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2521 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     2581 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     2641 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     2701 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     2761 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     2821 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2881 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2941 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     3001 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     3061 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     3121 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     3181 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     3241 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     3301 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     3361 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     3421 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     3481 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     3541 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca
     3601 ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     3661 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3721 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3781 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     3841 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     3901 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3961 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4021 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4081 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4141 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4201 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4261 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4321 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4381 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4441 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4501 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4561 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4621 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4681 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4741 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4801 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4861 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4921 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4981 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5041 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5101 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5161 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5221 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5281 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5341 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5401 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5461 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5521 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5581 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5641 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5701 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5761 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5821 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5881 ctgac
//
