pET30a-His-AviTag-SYT1-I368T(codon optimized)

LOCUS      pET30a-His-AviTag-SYT1-I368T(codon optimized)               6259 bp DNA     circular SYN 16-MAR-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6259)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 27, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(824..1279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     terminator      complement(1316..1363)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1416..2462
                     /label=His-AviTag-SYT1 I368T
     CDS             complement(2367..2411)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(2418..2435)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2445..2462)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2482..2504
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2519..2543
                     /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)."
     promoter        complement(2544..2562)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2875..2952
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2953..4035
                     /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"
     CDS             4844..5035
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    5137..5279
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5465..6053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             join(6175..6259,1..731)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
       61 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
      121 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
      181 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg
      241 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
      301 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
      361 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
      421 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
      481 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
      541 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
      601 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
      661 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
      721 agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa
      781 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat
      841 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
      901 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
      961 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1021 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     1081 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     1141 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     1201 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     1261 gcgccgctac agggcgcgtc ccattcgcca atccggatat agttcctcct ttcagcaaaa
     1321 aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg
     1381 cagcagccaa ctcagcttcc tttcgggctt tgtcattact ttttcaccgc taacatggcg
     1441 tcaacttctt cttccacttg aagggtgtgc cactgggcaa tcggacggcg cgggttcgcc
     1501 agcatgtctg accaatgacg taactccgcg ccggtgctgt tatagcccac gaacacctta
     1561 ccaattgcat cgtttttacc ggttttgtcg tagtccagaa ccgttaccac aacctggact
     1621 ttttgaatct gctcaaatgg aacctcgaac gagaagctct cattgtaata cggattcagg
     1681 gtgttctttt taatagtcgt cttctttttc ttcaggcgct taccgttctg catcaggtgg
     1741 attttaacgt acggatcaga gagaccgcca acatccatct ttttcaggtt ttttgcttcc
     1801 agaataacaa cggtcaactt acccgccgtc gggacgtatc tcaggctaaa gcagatatca
     1861 cccagttttt cttgctcctc tttctccgcg ctctgcagat cacgccactc ttcggtaaca
     1921 tggccgaaat cgacggtatt catcggtacc ttaaattcac cgatgatatc gtgtttgcta
     1981 aaacgatcga agtcataaac tgccatgact agggtcttgc cgcccaattc ggagtacggc
     2041 actttaaagg taaattgctc gttgaaaacc gggttcaaag tcttacgatg cactttggtt
     2101 tcgaacttct tcttcttgtc tgggagcagg aacactttca cgtacgggtc agaggtccca
     2161 cccatgtcca gagccggcag ttccgcagct tggatgatac ccacaagcag ctgattattc
     2221 tggaagtcat aatccaggct atattgcaac ttgcccaact tctcctcttc tttcggctcc
     2281 tctttctctt cgccatctgt caggcccgtc tcggcatcgt cgtccttcag cgcctggtct
     2341 ttcatagcgc taccgccacc agaaccttcg tgccattcga ttttctgcgc ctcaaaaatg
     2401 tcgttcaggc ccatgtggga accgcgcgga accaaaccgg aagaatggtg gtggtggtga
     2461 tgacttgatc ccatatgtat atctccttct taaagttaaa caaaattatt tctagagggg
     2521 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat
     2581 cgatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     2641 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     2701 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     2761 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     2821 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
     2881 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     2941 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     3001 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     3061 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     3121 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     3181 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     3241 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     3301 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     3361 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     3421 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     3481 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     3541 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     3601 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     3661 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     3721 attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc
     3781 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     3841 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     3901 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     3961 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     4021 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
     4081 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     4141 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     4201 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     4261 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     4321 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccccacg
     4381 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     4441 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     4501 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     4561 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     4621 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     4681 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     4741 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     4801 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
     4861 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
     4921 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
     4981 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5041 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5101 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     5161 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     5221 caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     5281 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5341 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5401 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5461 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5521 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5581 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5641 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5701 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5761 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5821 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5881 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5941 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6001 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6061 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6121 tggtcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     6181 catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat
     6241 ttatatgggt ataaatggg
//