LOCUS       sequence-455387-        7146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty 10xHis-SUMO (Smt3) fused Avi-tag, HRV-3C cleavable vector for 
            rapid cloning/DNA assembly via SwaI restriction site for E. coli 
            based protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7146)
  TITLE     Molecular Toolbox
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7146)
  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: 
            "Unpublished"
COMMENT     Sequence Label: MZB205
FEATURES             Location/Qualifiers
     source          1..7146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1002..1021
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1041..1118)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1324..1343
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1427..1446
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1427..1445
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1446..1470
                     /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             1485..1507
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1521..1550
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1851..1895
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1899..1922
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1966..1983
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2036..2054)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2050..2097
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2134..2589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2221..2240)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2431..2452
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2615..2719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2720..2788,2789..3577)
                     /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
                     LIKH"
     primer_bind     complement(2938..2957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5089..5677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5578..5597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5831..5848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5863..6002
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5928..5947)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6088..6110
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6104..6295)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7104..7146,1..1040))
                     /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"
ORIGIN
        1 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
       61 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
      121 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
      181 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
      241 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
      301 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
      361 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
      421 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
      481 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
      541 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
      601 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
      661 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
      721 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
      781 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
      841 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
      901 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
      961 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     1021 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     1081 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     1141 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     1201 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     1261 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     1321 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     1381 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
     1441 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
     1501 aaggagatat acatatgggc catcaccatc accatcacca tcaccatcac ggcagcatga
     1561 gcgatagcga agtgaaccag gaagcgaaac cggaagtgaa accggaagtg aaaccggaaa
     1621 cccatattaa cctgaaagtg agcgatggca gcagcgaaat tttttttaaa attaaaaaaa
     1681 ccaccccgct gcgtcgtctg atggaagcgt ttgcgaaacg tcagggcaaa gaaatggata
     1741 gcctgcgttt tctgtatgat ggcattcgta ttcaggcgga tcagaccccg gaagatctgg
     1801 atatggaaga taacgatatt attgaagcgc atcgtgaaca gattggcggc ggcctgaacg
     1861 atatttttga agcgcagaaa attgaatggc atgaaggtct ggaagtgctg tttcagggtc
     1921 cgggtagcgg catttaaata ataagcttcc atgggtaccc tcgagcacca ccaccaccac
     1981 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2041 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2101 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     2161 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2221 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2281 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2341 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2401 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2461 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2521 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     2581 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2641 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2701 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2761 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2821 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2881 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2941 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3001 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3061 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3121 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3181 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3241 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3301 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3361 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3421 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3481 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3541 gatcgctgag ataggtgcct cactgattaa gcactgagag atcccctcat aatttcccca
     3601 aaacgtaacc atgtgtgaat agattttgag taagcagggt tgcagccacg agtgagtctt
     3661 cccttgttat tgtgtagcca gaatgccgca aaacttccat gcctaagcga actgttgaga
     3721 gtacgtttcg atttctgact gtgttagcct ggaagtgctt gtcccaacct tgtttctgag
     3781 catgaacgcc cgcaagccaa catgttagtt gaagcatcag ggcgattagc agcatgatat
     3841 caaaacgctc tgagctgctc gttcggctat ggcgtaggcc tagtccgtag gcaggacttt
     3901 tcaagtctcg gaaggtttct tcaatctgca ttcgcttcga atagatatta acaagttgtt
     3961 tgggtgttcg aatttcaaca ggtaagttag ttgctagaat ccatggctcc tttgccgacg
     4021 ctgagtagat tttaggtgac gggtggtgac aatgagtccg tgtcgagcgc tgattttttc
     4081 ggcctttaga gcgagattta tacaatagaa tttggcatga gattgggttg cttttagtca
     4141 gcctcttata gcctaaagtc tttgagtgac tagatgacat atcatgtaag ttgctgatag
     4201 gtttccagtt ttccgctcct aggtctgcat attgtacttt tcctcttact cgacttaacc
     4261 agtaccaacc cagcttctca acggatttat accatggcac tttaaagcca gcatcactga
     4321 caatgagcgg tgtggtgtta ctcggtagaa tgctcgcaag gtcggctaga aattggtcat
     4381 gagctttctt tgaacattgc tctgaaagcg ggaacgcttt ctcataaaga gtaacagaac
     4441 gaccgtgtag tgcgactgaa gctcgcaata ccataagccg tttttgctca cggatatcag
     4501 accagtcaac aagtacaatg ggcatcgtat tgcccgaaca gataaagcta gcatgccaac
     4561 ggtatacagc gagtcgctct ttgtggaggt gacgattacc taacaatcgg tcgattcgtt
     4621 tgatgttatg ttttgttctc gctttggttg gcaggttacg gccaagttcg gtaagagtga
     4681 gagttttaca gtcaagtaag gcgtggcaag ccaacgttaa gctgttgagt cgttttaagt
     4741 gtaattcggg gcagaattgg taaagagagt cgtgtaaaat atcgagttcg cacattttgt
     4801 tgtctgatta ttgatttttg gcgaaaccat ttgatcatat gacaagatgt gtatctacct
     4861 taacttaatg attttgataa aaatcattag gggattcatc aggattaagc attggtaact
     4921 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5221 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5701 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5761 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5821 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     5881 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     5941 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     6001 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     6061 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     6121 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     6181 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     6241 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     6301 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     6361 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     6421 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     6481 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     6541 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     6601 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     6661 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     6721 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     6781 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     6841 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     6901 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     6961 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     7021 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     7081 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7141 agctgc
//
