LOCUS       Exported                4837 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 4837)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Apr 25, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4837
                     /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=Biotin
     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..1341
                     /label=New oligo
     CDS             1502..2035
                     /codon_start=1
                     /label=GentR
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     misc_feature    1502..1523
                     /label=oligo 5349
                     /note="Gent with h142, for SOEing."
     misc_feature    complement(1789..1809)
                     /label=oligo 1216
     misc_feature    1817..1835
                     /label=oligo1215
     misc_feature    1817..1834
                     /label=oligo 1222
     misc_feature    complement(2014..2035)
                     /label=oligo 5352
                     /note="Gent with h142,for SOEing"
     misc_feature    complement(2017..2041)
                     /label=oligo 2214
                     /note="PromGentRvNsi"
     misc_feature    join(2036..4837,1..2035)
                     /label=Untitled
     misc_feature    complement(2155..2182)
                     /label=New oligo
     misc_feature    2172..2177
                     /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    2178..2248
                     /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    2186..3043
                     /label=OriT
     misc_feature    complement(2275..2297)
                     /label=oligo 2831
                     /note="reverse oligo for OriT ( CV vector)"
     misc_feature    complement(2401..2428)
                     /label=oligo 4423
                     /note="reverse oligo for OriT ( CV vector)"
     misc_feature    complement(3030..3043)
                     /label=2146
     misc_feature    3079..3921
                     /label=AmpR
     misc_feature    3914..4302
                     /label=pT7pHip-OriT
                     /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    3975..4837
                     /label=OriC
     misc_feature    4680..4708
                     /label=oligo 4294
                     /note="For sequencing"
     misc_feature    4836..4875
                     /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 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     1381 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttgtacagtc tatgcctcgg
     1441 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     1501 gatgttacgc agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg
     1561 tggctcaagt atgggcatca ttcgcacatg taggctcggc cctgaccaag tcaaatccat
     1621 gcgggctgct cttgatcttt tcggtcgtga gttcggagac gtagccacct actcccaaca
     1681 tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattca tcgcgcttgc
     1741 tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaggtttga
     1801 gcagccgcgg agtgaaatct atatctatga tctcgcagtc tccggcgagc accggaggca
     1861 gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta
     1921 tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt
     1981 gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca cctaaatgca
     2041 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     2101 ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa
     2161 aatcaccaac ttctagaagc ttctgcctgg cggccttttg cgcctgatgc ggtattttct
     2221 ccttacgcat ctgtgcggta tttcacaccg catatcgaca tccgccctca ccgccaggaa
     2281 cgcaaccgca gcctcatcac gccggcgctt cttggccgcg cgggattcaa cccactcggc
     2341 cagctcgtcg gtgtagctct ttggtatcgt ctctcgcctg tcccctcagt tcagtaattt
     2401 cctgcatttg cctgtttcca gtcggtagat attccacaaa acagcaggga agcagcgctt
     2461 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     2521 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     2581 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     2641 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     2701 ggctaccgcc ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag
     2761 gaagggcagc ccacctatca aggtgtactg ccttccagac gaacgaagag cgattgagga
     2821 aaaggcggcg gcggccggca tgagcctgtc ggcctacctg ctggccgtcg gccagggcta
     2881 caaaatcacg ggcgtcgtgg actatgagca cgtccgcgag ctggcccgca tcaatggcga
     2941 cctgggccgc ctgggcggcc tgctgaaact ctggctcacc gacgacccgc gcacggcgcg
     3001 gttcggtgat gccacgatcc tcgccctgct ggcgaagatc gactctagct agacctgcag
     3061 gattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3121 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3181 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3241 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3301 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3361 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3421 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3481 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3541 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3601 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3661 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3721 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3781 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3841 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3901 atcgctgaga taggtgcctc actgattaag cattggtaac cctgcaggat gcatttaaaa
     3961 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4021 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4081 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4141 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4201 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4261 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4321 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4381 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4441 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4501 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4561 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4621 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4681 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4741 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4801 ccgagcgcag cgagtcagtg agcgaggaag cggaaga
//
