LOCUS       Exported                5128 bp ds-DNA     circular SYN 27-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5128)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Apr 25, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5128
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..16
                     /label=untitled 2
     misc_feature    complement(17..40)
                     /label=oligo 2176
     misc_feature    39..44
                     /label=rbs
     CDS             52..1320
                     /codon_start=1
                     /label=polyhisbioMbpTEV
                     /translation="MHHHHHHHHGSLNDIFEAQKIEWHEYDIPTTENLKTEEGKLVIWI
                     NGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGY
                     AQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTW
                     EEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGA
                     KAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVL
                     PTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKS
                     YEEELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQ
                     TNSSSNNNNNNNNNNENLYFQGS"
     misc_feature    55..78
                     /label=8 His
     misc_feature    complement(79..120)
                     /label=Biotinylatiion sequence
     misc_feature    complement(103..120)
                     /label=oligo 5217 or oligo 5272
                     /note="with BamHI site."
     misc_feature    complement(106..153)
                     /label=oligo 4425
     misc_feature    121..168
                     /label=oligo 4424
     misc_feature    496..522
                     /label=2492
                     /note="Forward primer designed to replace a G to an A, 
                     thereby destroying the bglII site
                     "
     misc_feature    complement(498..525)
                     /label=2493
                     /note="Reverse primer designed to replace a G with an T, 
                     thereby destroying the bglII site.
                     "
     misc_feature    1210..1233
                     /label=Mal E Sequencing Primer
     misc_feature    1212..1234
                     /label=o577
                     /note="Oligo 577"
     misc_feature    1212..1233
                     /label=pMalc25' oligo binding site
                     /note="Oligo on order"
     misc_feature    1212..1230
                     /label=pMalc25' oligo binding site
                     /note="Oligo on order
                     "
     misc_feature    complement(1275..1291)
                     /label=1528
                     /note="Reverse oligo designed to introduce TEV protease 
                     cleavage site after the polyglutamine region in the pMal 
                     vectors."
     misc_feature    1294..1309
                     /label=1299
     misc_feature    1312..1344
                     
     misc_feature    1326..1345
                     /label=oligo 1014
     misc_feature    1359..1364
                     /label=-35_signal
                     /note="     -35_signal      94..99
                                          /label=-35
                                          /SECDrawAs=""Label""
                                          /SECName=""-35"""
     misc_feature    1379..1387
                     /label=-10_signal
                     /note="     -10_signal      114..122
                                          /label=-10
                                          /SECDrawAs=""Label""
                                          /SECName=""-10"""
     misc_feature    1435..1449
                     /label=oligo 532
     CDS             1438..2253
                     /codon_start=1
                     /label=KanR
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    1438..1454
                     /label=oligo 148
     misc_feature    1451..1470
                     /label=OLIGO 019 (Tn903A)
     misc_feature    complement(2133..2152)
                     /label=OLIGO 020 (TN903B)
     misc_feature    complement(2232..2257)
                     /label=oligo 983
     misc_feature    complement(2242..2259)
                     /label=oligo 149
     misc_feature    complement(2245..2255)
                     /label=oligo 533
     misc_feature    complement(2445..2473)
                     
     misc_feature    2463..2468
                     /label=e58 [168-1309]
                     /note="e058.e1834. pUK4K plasmid. The plasmid contains a 
                     kanR gene block from Tn903 inserted between MCS within lacZ
                     gene (N-terminal region)."
     misc_feature    2469..2539
                     /label=C.V.-T7HIp Tat SufE No tag
                     /note="This file was downloaded from the 12/87 VecBase by 
                     Keith A Johnson on Dec. 14, 1989.
                     Modified by Bob Gross, 12/26/89. 
                     ENTRY BLUEKSM                  #TYPE DNA CIRCULAR
                     TITLE BlueScribe KS Minus - Cloning vector
                     DATE    28-JAN-1987
                        #sequence  02-FEB-1987
                        #sequence  04-MAR-1987
                        #sequence  03-APR-1987
                     ACCESSION VB0077
                     SOURCE    artificial
                     REFERENCE
                        #number 1
                        #authors Fernandez J.M., Short J.M., Renshaw M., Huse 
                     W., Sorge J.
                        #journal Gene (1987) in press
                        #citation Sequence data from StrataGene
                        #comment  sequence correction according to Stratagene
                     COMMENT
                        Obtained from StrataGene on floppy disc.
                        Revised 02-FEB-1987 by F. Pfeiffer:
                        1409/10 ""AT"" to ""TA"" to match revised sequence of 
                     PBR322
                        Revised 4-MAR-1987 to match sequence of pUC19 on request
                        of Stratagene
                        Polylinker region revised 03-APR-1987 according to 
                     Stratagene
                     COMMENT
                        The stand shown corresponds to pUC19c.
                        As in the published sequence of pUC19c, The M13mp19 lacZ
                     region
                        is on the complementary strand.
                     COMMENT
                        This vector contains the f1 origin so that the minus 
                     strand
                        can be obtained upon f1 superinfection.
                     KEYWORDS
                     CROSSREFERENCE
                        #parent
                          VecBase(3):BlueM13m
                        #parent
                          VecBase(3):pUC19, VecSource(3):PromT7, 
                     VecSource(3):PromT3,
                          VecSource(3):bGalKS, GenBank(50):PF1
                        #brother
                          VecBase(3):BlueKSp, VecBase(3):BlueSKm
                     PARENT
                        Features of BlueKSm (2964 bp)
                          residue     source
                           3- 458  5488-5943     phage f1
                         460- 624   236- 400     pUC19c
                         626- 645     1-  20     T7 promoter
                         653- 760   108-   1 (c) BlueKS-polylinker
                         772- 791    20-   1 (c) T3 promoter
                         795-2964   448-2617     pUC19c
                        Conflict (cfl) and Mutations (mut): none
                     PARENT
                        Features of BlueKSm (2964 bp)
                          residue     source
                           3- 458  5488-5943     phage f1
                         460- 624   449- 285 (c) pUC19
                         626- 645     1-  20     T7 promoter
                         653- 760   108-   1 (c) BlueKS-polylinker
                         772- 791    20-   1 (c) T3 promoter
                         795-1031   237-   1 (c) pUC19
                        1032-2964  2686- 754 (c) pUC19
                        Conflict (cfl) and Mutations (mut): none
                     FEATURE
                              643           start of T7-RNA synthesis
                              774       (c) start of T3-RNA synthesis
                        1976-2764 789-1 (c) Ap-R; b-lactamase
                     POLYLINKER
                          
                     KpnI-DraII-ApaI-XhoI-SalI-ClaI-HindIII-EcoRV-EcoRI-PstI-
                          SmaI-BamHI-SpeI-XbaI-NotI-XmaIII-BstXI-SacII-SacI
                     SELECTION
                        #resistance  Ap
                        #indicator beta-galactosidase
                     SUMMARY  BlueKSm    #length 2964   #checksum 1589
                     SEQUENCE
                     Vecbase:Blueksm  Length: 2964  December 11, 1989  10:03  
                     Check: 1589  ..P"
     misc_feature    2477..3334
                     /label=OriT
     misc_feature    complement(2566..2588)
                     /label=oligo 2831
                     /note="reverse oligo for OriT ( CV vector)"
     misc_feature    complement(2692..2719)
                     /label=oligo 4423
                     /note="reverse oligo for OriT ( CV vector)"
     misc_feature    complement(3321..3334)
                     /label=2146
     misc_feature    3370..4212
                     /label=AmpR
     misc_feature    4266..5128
                     /label=OriC
     misc_feature    4971..4999
                     /label=oligo 4294
                     /note="For sequencing"
     misc_feature    5127..5166
                     /label=oligo 4428
                     /note="with EcoRI site."
ORIGIN
        1 gcgcggccgc ttaatacgac tcactatagg gagaattcaa ggagctttca tatgcaccat
       61 catcatcacc atcaccatgg atctctgaac gacatcttcg aagctcagaa aatcgaatgg
      121 catgaatacg atatcccaac gaccgaaaac ctgaaaactg aagaaggtaa actggtaatc
      181 tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa
      241 gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag
      301 gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc
      361 tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg
      421 tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct
      481 gttgaagcgt tatcgctgat ttataacaaa gatttgctgc cgaacccgcc aaaaacctgg
      541 gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc
      601 aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc
      661 aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa
      721 gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat
      781 tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg
      841 tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc
      901 ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc
      961 agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac tgatgaaggt
     1021 ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa
     1081 gagttggcga aagatccacg tattgccgcc accatggaaa acgcccagaa aggtgaaatc
     1141 atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac
     1201 gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac taattcgagc
     1261 tcgaacaaca acaacaataa caataacaac aacgaaaacc tgtattttca gggatcctaa
     1321 gggggggaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     1381 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     1441 agccatattc aacgggaaac gtcttgctcg aggccgcgat taaattccaa catggatgct
     1501 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1561 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1621 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1681 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1741 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1801 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1861 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1921 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1981 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2041 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2101 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2161 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2221 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2281 tggcagagca ttacgctgac ttgacgggac ggcggctttg ttgaataaat cgaacttttg
     2341 ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc
     2401 aaaagttcaa aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc
     2461 tctctagaag cttctgcctg gcggcctttt gcgcctgatg cggtattttc tccttacgca
     2521 tctgtgcggt atttcacacc gcatatcgac atccgccctc accgccagga acgcaaccgc
     2581 agcctcatca cgccggcgct tcttggccgc gcgggattca acccactcgg ccagctcgtc
     2641 ggtgtagctc tttggtatcg tctctcgcct gtcccctcag ttcagtaatt tcctgcattt
     2701 gcctgtttcc agtcggtaga tattccacaa aacagcaggg aagcagcgct tttccgctgc
     2761 ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     2821 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     2881 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     2941 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cggctaccgc
     3001 cggcgtaaca gatgagggca agcggatggc tgatgaaacc aagccaacca ggaagggcag
     3061 cccacctatc aaggtgtact gccttccaga cgaacgaaga gcgattgagg aaaaggcggc
     3121 ggcggccggc atgagcctgt cggcctacct gctggccgtc ggccagggct acaaaatcac
     3181 gggcgtcgtg gactatgagc acgtccgcga gctggcccgc atcaatggcg acctgggccg
     3241 cctgggcggc ctgctgaaac tctggctcac cgacgacccg cgcacggcgc ggttcggtga
     3301 tgccacgatc ctcgccctgc tggcgaagat cgactctagc tagacctgca ggattgaaaa
     3361 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3421 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3481 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3541 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3601 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3661 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3721 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3781 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3841 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3901 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3961 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4021 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4081 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4141 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4201 ataggtgcct cactgattaa gcattggtaa ccctgcagga tgcatttaaa aggatctagg
     4261 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4321 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4381 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4441 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4501 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4561 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4621 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4681 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4741 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4801 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4861 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4921 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4981 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5041 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5101 gcgagtcagt gagcgaggaa gcggaaga
//
