LOCUS       Exported                6193 bp ds-DNA     circular SYN 22-OCT-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6193)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 11, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6193
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        22..38
                     /label=T7 promoter
                     /label=T7\promoter
                     /note="vntifkey=30"
     CDS             41..65
                     /codon_start=1
                     /label=lac operator
                     /label=lac\operator
                     /note="vntifkey=4"
                     /translation="GIVSG*QF"
     CDS             109..120
                     /codon_start=1
                     /label=Start seq
                     /translation="MGSS"
     misc_binding    121..138
                     /label=His-tag
                     /note="vntifkey=20"
     misc_binding    148..165
                     /label=thrombin site
                     /label=thrombin\site
                     /note="vntifkey=20"
     CDS             169..411
                     /codon_start=1
                     /label=SUMO1
                     /note="modified SUMO1, added restr. sites via silent 
                     mutation
                     vntifkey=4"
                     /translation="MEGEYIKLKVIGQDSSEIHFKVKMTTHLKKLKESYCQRQGVPMNS
                     LRFLFEGQRIADNHTPKELGMEEEDVIEVYQEQTGG"
     primer_bind     complement(390..431)
                     /label=Vector.REV
     primer_bind     392..432
                     /label=Fragment.FOR
     CDS             412..1050
                     /codon_start=1
                     /label=CHMP2B
                     /translation="MASLFKKKTVDDVIKEQNRELRGTQRAIIRDRAALEKQEKQLELE
                     IKKMAKIGNKEACKVLAKQLVHLRKQKTRTFAVSSKVTSMSTQTKVMNSQMKMAGAMST
                     TAKTMQAVNKKMDPQKTLQTMQNFQKENMKMEMTEEMINDTLDDIFDGSDDEEESQDIV
                     NQVLDEIGIEISGKMAKAPSAARSLPSASTSKATISDEEIERQLKALGVD"
     primer_bind     complement(888..923)
                     /label=Primer 2
     primer_bind     888..923
                     /label=Primer 1
     misc_feature    904
                     /label=T instead of C for 165 DELTA
     primer_bind     complement(1030..1072)
                     /label=Fragment.REV
     primer_bind     1031..1072
                     /label=Vector.FOR
     protein_bind    1075..1092
                     /label=His tag
                     /label=His\tag
                     /note="vntifkey=31"
     misc_feature    1093..1095
                     /label=Stop
                     /note="vntifkey=21"
     terminator      1160..1206
                     /label=T7 terminator
                     /label=T7\terminator
                     /note="vntifkey=43"
     rep_origin      1243..1698
                     /label=f1 origin
                     /label=f1\origin
                     /note="vntifkey=33"
     CDS             complement(1794..2606)
                     /codon_start=1
                     /label=kan sequence
                     /label=kan\sequence
                     /note="vntifkey=4"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3315
                     /label=ColE1 pBR322 origin
                     /label=ColE1\pBR322\origin
                     /note="vntifkey=33"
     CDS             complement(4749..5828)
                     /codon_start=1
                     /label=lac I
                     /label=lac\I
                     /note="vntifkey=4"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
       61 attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat gggcagcagc
      121 catcatcatc atcatcacag cagcggcctg gtgccgcgcg gcagccatat ggaaggtgaa
      181 tatattaaac tcaaagtcat tggacaggat agcagtgaga ttcacttcaa agtgaaaatg
      241 acaacacatc tcaagaaact caaagaatca tactgtcaaa gacagggtgt tccaatgaat
      301 tcactcaggt ttctctttga gggtcagaga attgctgata atcatactcc aaaagaactg
      361 ggaatggagg aagaagatgt gattgaagtc taccaggaac aaaccggtgg aatggcgtcc
      421 ctcttcaaga agaaaaccgt ggatgatgta ataaaggaac agaatcgaga gttacgaggt
      481 acacagaggg ctataatcag agatcgagca gctttagaga aacaagaaaa acagctggaa
      541 ttagaaatta agaaaatggc caagattggt aataaggaag cttgcaaagt tttagccaaa
      601 caacttgtgc atctacggaa acagaagacg agaacttttg ctgtaagttc aaaagttact
      661 tctatgtcta cacaaacaaa agtgatgaat tcccaaatga agatggctgg agcaatgtct
      721 accacagcaa aaacaatgca ggcagttaac aagaagatgg atccacaaaa gacattacaa
      781 acaatgcaga atttccagaa ggaaaacatg aaaatggaaa tgactgaaga aatgatcaat
      841 gatacacttg atgacatctt tgacggttct gatgacgaag aagaaagcca ggatattgtg
      901 aatcaagttc ttgatgaaat tggaattgaa atttctggaa agatggccaa agctccatca
      961 gctgctcgaa gcttaccatc tgcctctact tcaaaggcta caatctcaga tgaagagatt
     1021 gaacggcaac tcaaggcttt aggagtagat tgaaagcttg cggccgcact cgagcaccac
     1081 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     1141 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1201 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     1261 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1321 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1381 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1441 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1501 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1561 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     1621 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1681 tattaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1741 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     1801 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1861 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     1921 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     1981 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     2041 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     2101 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     2161 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     2221 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     2281 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     2341 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     2401 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     2461 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     2521 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     2581 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     2641 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2701 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2761 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2821 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2881 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2941 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3001 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3061 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3121 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3181 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3241 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3301 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3361 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3421 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3481 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3541 ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3601 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3661 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3721 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3781 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3841 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3901 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     3961 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     4021 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     4081 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     4141 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     4201 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     4261 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     4321 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     4381 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     4441 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     4501 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     4561 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     4621 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     4681 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4741 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4801 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4861 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     4921 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     4981 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     5041 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     5101 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     5161 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     5221 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     5281 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     5341 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5401 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     5461 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     5521 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     5581 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     5641 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     5701 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     5761 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     5821 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     5881 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     5941 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     6001 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     6061 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     6121 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     6181 ggcgtagagg atc
//
