LOCUS       sequence-458447-        5727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty 10xHis-SUMO (Smt3) 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 5727)
  TITLE     Molecular Toolbox
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5727)
  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            "Unpublished"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: MZB184
FEATURES             Location/Qualifiers
     source          1..5727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1582..1773
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1767..1789)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1875..2014
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1930..1949
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2029..2046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2200..2788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2280..2299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2959..3750,3751..3819))
                     /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     3582..3601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3820..3924)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3950..4405)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4087..4108)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4299..4318
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4442..4489)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4485..4503
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4556..4573)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4908..4937)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             4951..4973
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4988..5012
                     /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)."
     primer_bind     complement(5012..5031)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5013..5031)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5115..5134)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5340..5417
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5418..5727,1..773)
                     /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     complement(5437..5456)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
       61 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
      121 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
      181 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
      241 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
      301 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
      361 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
      421 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
      481 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
      541 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
      601 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
      661 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
      721 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
      781 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
      841 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
      901 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
      961 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     1021 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     1081 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
     1141 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     1201 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     1261 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     1321 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     1381 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     1441 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     1501 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     1561 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     1621 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     1681 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     1741 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     1801 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     1861 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     1921 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattgcggt gtgaaatacc
     1981 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     2041 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2101 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2161 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2221 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2281 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2401 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2521 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2581 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2641 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2701 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2761 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2821 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2881 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2941 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3001 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3061 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3241 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
     3301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3361 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3421 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     3961 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4021 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4081 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4141 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4201 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4261 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4321 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4381 cttaatgcgc cgctacaggg cgcgtcccat tcgccaatcc ggatatagtt cctcctttca
     4441 gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag
     4501 cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tctcagtggt
     4561 ggtggtggtg gtgctcgagg gtacccatgg atccttatta tttaaatgcc gccaatctgt
     4621 tcacgatgcg cttcaataat atcgttatct tccatatcca gatcttccgg ggtctgatcc
     4681 gcctgaatac gaatgccatc atacagaaaa cgcaggctat ccatttcttt gccctgacgt
     4741 ttcgcaaacg cttccatcag acgacgcagc ggggtggttt ttttaatttt aaaaaaaatt
     4801 tcgctgctgc catcgctcac tttcaggtta atatgggttt ccggtttcac ttccggtttc
     4861 acttccggtt tcgcttcctg gttcacttcg ctatcgctca tgctgccgtg atggtgatgg
     4921 tgatggtgat ggtgatggcc catatgtata tctccttctt aaagttaaac aaaattattt
     4981 ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg
     5041 gatcgagatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     5101 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     5161 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     5221 gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     5281 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gagatcccgg
     5341 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     5401 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     5461 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     5521 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     5581 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     5641 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     5701 tggtgtcgat ggtagaacga agcggcg
//
