LOCUS       sequence-455379-        5775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty 10xHis-SUMO (Smt3) fused Avi-tag 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 5775)
  TITLE     Molecular Toolbox
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5775)
  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            "Unpublished"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: MZB186
FEATURES             Location/Qualifiers
     source          1..5775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        135..239
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(240..308,309..1100)
                     /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     complement(458..477)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1271..1859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1760..1779
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2013..2030
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2045..2184
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2110..2129)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2270..2292
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2286..2477)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3286..4368)
                     /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     4330..4349
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4369..4446)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4652..4671
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4755..4774
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4755..4773
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4774..4798
                     /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             4813..4835
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4849..4878
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5179..5223
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5261..5278
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5331..5349)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5345..5392
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      join(5429..5775,1..109)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5516..5535)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5726..5747
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
       61 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
      121 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      181 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      241 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
      301 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
      361 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
      421 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      481 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      541 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      601 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      661 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      721 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      781 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      841 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      901 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      961 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     1021 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     1081 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1141 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1201 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1261 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1321 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1381 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     1441 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1501 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1561 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1621 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1681 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1741 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1801 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1861 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1921 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1981 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     2041 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     2101 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     2161 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     2221 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     2281 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     2341 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     2401 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     2461 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     2521 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     2581 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     2641 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     2701 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     2761 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     2821 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     2881 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     2941 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     3001 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     3061 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     3121 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     3181 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     3241 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     3301 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3361 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     3421 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     3481 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     3541 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     3601 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     3661 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     3721 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     3781 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     3841 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     3901 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     3961 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     4021 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     4081 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     4141 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     4201 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     4261 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     4321 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     4381 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     4441 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     4501 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     4561 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     4621 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     4681 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     4741 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     4801 tctagaaata attttgttta actttaagaa ggagatatac atatgggcca tcaccatcac
     4861 catcaccatc accatcacgg cagcatgagc gatagcgaag tgaaccagga agcgaaaccg
     4921 gaagtgaaac cggaagtgaa accggaaacc catattaacc tgaaagtgag cgatggcagc
     4981 agcgaaattt tttttaaaat taaaaaaacc accccgctgc gtcgtctgat ggaagcgttt
     5041 gcgaaacgtc agggcaaaga aatggatagc ctgcgttttc tgtatgatgg cattcgtatt
     5101 caggcggatc agaccccgga agatctggat atggaagata acgatattat tgaagcgcat
     5161 cgtgaacaga ttggcggcgg cctgaacgat atttttgaag cgcagaaaat tgaatggcat
     5221 gaaggtattt aaataataag cttccatggg taccctcgag caccaccacc accaccactg
     5281 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     5341 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     5401 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     5461 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5521 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5581 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5641 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5701 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5761 cctatctcgg tctat
//
