LOCUS       209115-attachmen        5743 bp DNA     circular SYN 21-DEC-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5743)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 21, 2023 from SnapGene 7.0.2
            https://www.snapgene.com
COMMENT     pET-28 a (+)
FEATURES             Location/Qualifiers
     source          1..5743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label=f1 ori
                     /note="direction: RIGHT note: f1 bacteriophage origin of 
                     replication; arrow indicates direction of (+) strand 
                     synthesis"
                     /note="color: #ffef86"
     CDS             complement(560..1375)
                     /label=KanR
                     /note="codon_start: 1 gene: aph(3')-Ia note: confers 
                     resistance to kanamycin in bacteria or G418 (Geneticin(R)) 
                     in eukaryotes product: aminoglycoside phosphotransferase 
                     transl_table: 1 translation: 
                     MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKG
                     SVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGE
                     NIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQ
                     VWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCL
                     GEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
                     /note="color: #b7e6d7; direction: LEFT"
     rep_origin      1497..2085
                     /label=ori
                     /note="direction: RIGHT note: high-copy-number 
                     ColE1/pMB1/pBR322/pUC origin of replication"
                     /note="color: #ffef86"
     misc_feature    2271..2413
                     /label=bom
                     /note="note: basis of mobility region from pBR322"
                     /note="color: #b4abac"
     CDS             complement(2515..2706)
                     /label=rop
                     /note="codon_start: 1 gene: rop product: Rop protein, which
                     maintains plasmids at low copy number transl_table: 1 
                     translation: 
                     VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDD
                     GENL*"
                     /note="color: #993366; direction: LEFT"
     CDS             complement(3515..4597)
                     /label=lacI
                     /note="codon_start: 1 gene: 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). product: lac
                     repressor transl_table: 1 translation: 
                     VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGK
                     QSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQR
                     VSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVAL
                     GHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEG
                     IVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRL
                     LGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVS
                     RLESGQ*"
                     /note="color: #993366; direction: LEFT"
     promoter        complement(4598..4675)
                     /label=lacI promoter
                     /note="gene: lacI note: "
                     /note="color: #c6c9d1; direction: LEFT"
     promoter        4984..5002
                     /label=T7 promoter
                     /note="note: promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #c6c9d1"
     protein_bind    5003..5027
                     /label=lac operator
                     /note="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)."
                     /note="color: #31849b"
     RBS             5042..5064
                     /label=RBS
                     /note="note: efficient ribosome binding site from 
                     bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #b4abac"
     CDS             5083..5100
                     /label=6xHis
                     /note="codon_start: 1 product: 6xHis affinity tag 
                     transl_table: 1 translation: HHHHHH"
                     /note="color: #ff9ccd; direction: RIGHT"
     CDS             5113..5406
                     /label=SUMO
                     /note="codon_start: 1 transl_table: 1 translation: 
                     GSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRRLMEAFAKRQGKE
                     MDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
                     /note="color: #993366; direction: RIGHT"
     misc_feature    5155
                     /label=Is mutated to G (hence = E)
                     /note="color: #b4abac"
     CDS             5407..5433
                     /label=Linker Ref
                     /label=Linker Ref.
                     /note="codon_start: 1 transl_table: 1 translation: 
                     SGSCSGSGS"
                     /note="color: #f58a5e; direction: RIGHT"
     CDS             5434..5445
                     /label=Consensus N-terminal
                     /note="codon_start: 1 transl_table: 1 translation: SPED"
                     /note="color: #ffef86; direction: RIGHT"
     CDS             5446..5529
                     /label=A' CC
                     /note="codon_start: 1 transl_table: 1 translation: 
                     KIAQLKQKIQALKQENQQLEEENAALEY"
                     /note="color: #f58a5e; direction: RIGHT"
     CDS             5539..5562
                     /label=Strep-Tag II
                     /note="codon_start: 1 product: peptide that binds 
                     Strep-Tactin(R), an engineered form of streptavidin 
                     transl_table: 1 translation: WSHPQFEK"
                     /note="color: #ff9ccd; direction: RIGHT"
     CDS             5587..5604
                     /label=6xHis
                     /note="codon_start: 1 product: 6xHis affinity tag 
                     transl_table: 1 translation: HHHHHH"
                     /note="color: #ff9ccd; direction: RIGHT"
     terminator      5671..5718
                     /label=T7 terminator
                     /note="note: transcription terminator for bacteriophage T7 
                     RNA polymerase"
                     /note="color: #c6c9d1"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     5041 ttttgtttaa ctttaagaag gagatatacc atgggcagca gtcatcatca tcatcatcac
     5101 agcagcggcg gtggtagcga tagcgaagtc aatcaagaag cgaaaccgga agtcgaaccg
     5161 gaagtcaaac cggaaacgca catcaacctg aaagttagtg atggcagctc tgaaattttc
     5221 tttaaaatta agaaaaccac gccgctgcgt cgcctgatgg aagcgtttgc caaacgccag
     5281 ggcaaagaaa tggatagcct gcgcttcctg tatgacggta ttcgtatcca ggcggatcaa
     5341 gccccggaag atctggacat ggaagataac gacattatcg aagcgcatcg cgaacagatt
     5401 ggcggcagtg gttcatgtag cggtagtgga tcctctccgg aagacaaaat tgcgcagttg
     5461 aaacaaaaaa ttcaggcgct caagcaagaa aatcaacagt tggaggagga gaatgcagcc
     5521 ctggagtatg gtggaagttg gtcccatccg cagttcgaga aataaaagct tgcggccgca
     5581 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     5641 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     5701 gtcttgaggg gttttttgct gaaaggagga actatatccg gat
//
