LOCUS       Exported                6848 bp ds-DNA     circular SYN 20-JAN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBad His6 MBP TEV LIC cloning vector (8C)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6848)
  TITLE     8-series E. coli arabinose expression vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6848)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 20, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6848
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      21..67
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     220..237
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(298..318)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(373..1473)
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     complement(376..399)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             complement(1483..1500)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1520..1542
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(1576..1860)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(1634..1653)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1887..2765
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(2924..2943)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             4401..4592
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4586..4608)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4694..4834
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4749..4768
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4849..4866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5020..5608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5100..5119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5778..6638)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6401..6420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6639..6743)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6811..6829
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 agttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
       61 ttatttgatg cctggcagtt ccctactctc gcatggggag accccacact accatcggcg
      121 ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg
      181 ccaggcaaat tctgttttat cagaccgctt ctgcgttctg atttaatctg tatcaggctg
      241 aaaatcttct ctcatccgcc aaaacagcca agggatccgt tatccacttc caatattgga
      301 ttggaagtac aggttttcct cgatcccgag gttgttgtta ttgttattgt tgttgttgtt
      361 cgagctcgaa ttagtctgcg cgtctttcag ggcttcatcg acagtctgac gaccgctggc
      421 ggcgttgatc accgcagtac gcacggcata ccagaaagcg gacatctgcg ggatgttcgg
      481 catgatttca cctttctggg cgttttccat agtggcggca atacgtggat ctttcgccaa
      541 ctcttcctcg taagacttca gcgctacggc acccagcggt ttgtctttat taaccgcttc
      601 cagaccttca tcagtcagca gatagttttc gaggaactct tttgccagct ctttgttcgg
      661 actggcggcg ttaatacctg cgctcagcac gccaacgaac ggtttggatg gttgaccctt
      721 gaaggtcggc agtaccgtta caccataatt cactttgctg gtgtcgatgt tggaccatgc
      781 ccacgggccg ttgatggtca tcgctgtttc gcctttatta aaggcagctt ctgcgatgga
      841 gtaatcggtg tctgcattca tgtgtttgtt tttaatcagg tcaaccagga aggtcagacc
      901 cgctttcgcg ccagcgttat ccacgcccac gtctttaatg tcgtacttgc cgttttcata
      961 cttgaacgca taacccccgt cagcagcaat cagcggccag gtgaagtacg gttcttgcag
     1021 gttgaacatc agcgcgctct tacctttcgc tttcagttct ttatccagcg ccgggatctc
     1081 ttcccaggtt tttggcgggt tcggcagcag atctttgtta taaatcagcg ataacgcttc
     1141 aacagcgatc gggtaagcaa tcagcttgcc gttgtaacgt acggcatccc aggtaaacgg
     1201 atacagcttg tcctggaacg ctttgtccgg ggtgatttca gccaacaggc cagattgagc
     1261 gtagccacca aagcggtcgt gtgcccagaa gataatgtca gggccatcgc cagttgccgc
     1321 aacctgtggg aatttctctt ccagtttatc cggatgctca acggtgactt taattccggt
     1381 atctttctcg aatttcttac cgacttcagc gagaccgtta tagcctttat cgccgttaat
     1441 ccagattacc agtttacctt cttcgatttt catagaagaa ccatggtgat ggtgatggtg
     1501 agaagatttc atatgtatat ctccttctta aagttaaaca aaattatttc tagatgttag
     1561 cccaaaaaaa cgggtatgga gaaacagtag agagttgcga taaaaagcgt caggtaagat
     1621 ccgctaatct tatggataaa aatgctatgg catagcaaag tgtgacgccg tgcaaataat
     1681 caatgtggac ttttctgccg tgattataga cacttttgtt acgcgttttt gtcatggctt
     1741 tggtcccgct ttgttacaga atgcttttaa taagcggggt taccggtttg gttagcgaga
     1801 agagccagta aaagacgcag tgacggcaat gtctgatgca atatggacaa ttggtttctt
     1861 ctctgaatgg cgggagtatg aaaagtatgg ctgaagcgca aaatgatccc ctgctgccgg
     1921 gatactcgtt taatgcccat ctggtggcgg gtttaacgcc gattgaggcc aacggttatc
     1981 tcgatttttt tatcgaccga ccgctgggaa tgaaaggtta tattctcaat ctcaccattc
     2041 gcggtcaggg ggtggtgaaa aatcagggac gagaatttgt ttgccgaccg ggtgatattt
     2101 tgctgttccc gccaggagag attcatcact acggtcgtca tccggaggct cgcgaatggt
     2161 atcaccagtg ggtttacttt cgtccgcgcg cctactggca tgaatggctt aactggccgt
     2221 caatctttgc caatacgggg ttctttcgcc cggatgaagc gcaccagccg catttcagcg
     2281 acctgtttgg gcaaatcatt aacgccgggc aaggggaagg gcgctattcg gagctgctgg
     2341 cgataaatct gcttgagcaa ttgttgctgc ggcgcatgga agcgattaac gagtcgctcc
     2401 atccaccgat ggataatcgg gtacgcgagg cttgtcagta catcagcgat cacctggcag
     2461 acagcaattt tgatatagcc agcgtcgcac agcatgtttg cttgtcgccg tcgcgtctgt
     2521 cacgtctttt ccgccagcag ttagggatta gcgtcttaag ctggcgcgag gaccaacgta
     2581 tcagccaggc gaagctgctt ttgagcacca cccggatgcc tatcgccacc gtcggtcgca
     2641 atgttggttt tgacgatcaa ctctatttct cgcgggtatt taaaaaatgc accggggcca
     2701 gcccgagcga gttccgtgcc ggttgtgaag aaaaagtgaa tgatgtagcc gtcaagttgt
     2761 cataattggt aacgaatcag acaattgacg gctcgaggga gtagcatagg gtttgcagaa
     2821 tccctgcttc gtccatttga cgggcacatt atggcggccg cgatcctcta cgccggacgc
     2881 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     2941 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     3001 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
     3061 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     3121 tcgcataagg gagagcgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     3181 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     3241 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     3301 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     3361 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     3421 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc
     3481 ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg
     3541 ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt
     3601 accagcctaa cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg
     3661 agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc
     3721 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
     3781 tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga
     3841 atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca
     3901 cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt
     3961 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
     4021 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
     4081 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
     4141 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
     4201 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
     4261 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
     4321 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
     4381 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     4441 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     4501 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     4561 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4621 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4681 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     4741 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     4801 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     4861 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4921 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4981 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5461 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5701 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5761 gtaaacttgg tcccgggtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5821 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5881 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5941 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6001 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6061 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     6121 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6181 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6241 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6301 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6361 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     6421 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6481 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6541 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6601 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatgttat
     6661 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6721 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6781 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     6841 caagaatt
//
