LOCUS       sequence-382470-        6906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6906)
  TITLE     8-series E. coli arabinose expression vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6906)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBAD His6 MBP TEV LIC transfer vector (8CT)
FEATURES             Location/Qualifiers
     source          1..6906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(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(327..347)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(402..1502)
                     /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(405..428)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             complement(1512..1529)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1549..1571
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(1634..1918)
                     /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(1692..1711)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1945..2823
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(2982..3001)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             4459..4650
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4644..4666)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4752..4892
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4807..4826
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4907..4924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5078..5666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5158..5177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5836..6627,6628..6696))
                     /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     6459..6478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6697..6801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6869..6887
                     /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 ggccggccac caggtggcgc gccgcgatcg
      301 cggatccgtt atccacttcc aatattggat tggaagtaca ggttttcctc gatcccgagg
      361 ttgttgttat tgttattgtt gttgttgttc gagctcgaat tagtctgcgc gtctttcagg
      421 gcttcatcga cagtctgacg accgctggcg gcgttgatca ccgcagtacg cacggcatac
      481 cagaaagcgg acatctgcgg gatgttcggc atgatttcac ctttctgggc gttttccata
      541 gtggcggcaa tacgtggatc tttcgccaac tcttcctcgt aagacttcag cgctacggca
      601 cccagcggtt tgtctttatt aaccgcttcc agaccttcat cagtcagcag atagttttcg
      661 aggaactctt ttgccagctc tttgttcgga ctggcggcgt taatacctgc gctcagcacg
      721 ccaacgaacg gtttggatgg ttgacccttg aaggtcggca gtaccgttac accataattc
      781 actttgctgg tgtcgatgtt ggaccatgcc cacgggccgt tgatggtcat cgctgtttcg
      841 cctttattaa aggcagcttc tgcgatggag taatcggtgt ctgcattcat gtgtttgttt
      901 ttaatcaggt caaccaggaa ggtcagaccc gctttcgcgc cagcgttatc cacgcccacg
      961 tctttaatgt cgtacttgcc gttttcatac ttgaacgcat aacccccgtc agcagcaatc
     1021 agcggccagg tgaagtacgg ttcttgcagg ttgaacatca gcgcgctctt acctttcgct
     1081 ttcagttctt tatccagcgc cgggatctct tcccaggttt ttggcgggtt cggcagcaga
     1141 tctttgttat aaatcagcga taacgcttca acagcgatcg ggtaagcaat cagcttgccg
     1201 ttgtaacgta cggcatccca ggtaaacgga tacagcttgt cctggaacgc tttgtccggg
     1261 gtgatttcag ccaacaggcc agattgagcg tagccaccaa agcggtcgtg tgcccagaag
     1321 ataatgtcag ggccatcgcc agttgccgca acctgtggga atttctcttc cagtttatcc
     1381 ggatgctcaa cggtgacttt aattccggta tctttctcga atttcttacc gacttcagcg
     1441 agaccgttat agcctttatc gccgttaatc cagattacca gtttaccttc ttcgattttc
     1501 atagaagaac catggtgatg gtgatggtga gaagatttca tatgtatatc tccttcttaa
     1561 agttaaacaa aattatttct agaactcgag tcctgcaggg cggccgctta attaatagcc
     1621 caaaaaaacg ggtatggaga aacagtagag agttgcgata aaaagcgtca ggtaagatcc
     1681 gctaatctta tggataaaaa tgctatggca tagcaaagtg tgacgccgtg caaataatca
     1741 atgtggactt ttctgccgtg attatagaca cttttgttac gcgtttttgt catggctttg
     1801 gtcccgcttt gttacagaat gcttttaata agcggggtta ccggtttggt tagcgagaag
     1861 agccagtaaa agacgcagtg acggcaatgt ctgatgcaat atggacaatt ggtttcttct
     1921 ctgaatggcg ggagtatgaa aagtatggct gaagcgcaaa atgatcccct gctgccggga
     1981 tactcgttta atgcccatct ggtggcgggt ttaacgccga ttgaggccaa cggttatctc
     2041 gattttttta tcgaccgacc gctgggaatg aaaggttata ttctcaatct caccattcgc
     2101 ggtcaggggg tggtgaaaaa tcagggacga gaatttgttt gccgaccggg tgatattttg
     2161 ctgttcccgc caggagagat tcatcactac ggtcgtcatc cggaggctcg cgaatggtat
     2221 caccagtggg tttactttcg tccgcgcgcc tactggcatg aatggcttaa ctggccgtca
     2281 atctttgcca atacggggtt ctttcgcccg gatgaagcgc accagccgca tttcagcgac
     2341 ctgtttgggc aaatcattaa cgccgggcaa ggggaagggc gctattcgga gctgctggcg
     2401 ataaatctgc ttgagcaatt gttgctgcgg cgcatggaag cgattaacga gtcgctccat
     2461 ccaccgatgg ataatcgggt acgcgaggct tgtcagtaca tcagcgatca cctggcagac
     2521 agcaattttg atatagccag cgtcgcacag catgtttgct tgtcgccgtc gcgtctgtca
     2581 cgtcttttcc gccagcagtt agggattagc gtcttaagct ggcgcgagga ccaacgtatc
     2641 agccaggcga agctgctttt gagcaccacc cggatgccta tcgccaccgt cggtcgcaat
     2701 gttggttttg acgatcaact ctatttctcg cgggtattta aaaaatgcac cggggccagc
     2761 ccgagcgagt tccgtgccgg ttgtgaagaa aaagtgaatg atgtagccgt caagttgtca
     2821 taattggtaa cgaatcagac aattgacggc tcgagggagt agcatagggt ttgcagaatc
     2881 cctgcttcgt ccatttgacg ggcacattat ggcggtcgcg atcctctacg ccggacgcat
     2941 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     3001 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
     3061 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
     3121 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     3181 gcataaggga gagcgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     3241 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     3301 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
     3361 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     3421 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     3481 ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt
     3541 ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct
     3601 gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac
     3661 cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg cctcggcgag
     3721 cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc
     3781 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     3841 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     3901 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     3961 cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     4021 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     4081 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     4141 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     4201 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     4261 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     4321 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     4381 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     4441 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     4501 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     4561 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     4621 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4681 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4741 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     4801 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     4861 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     4921 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4981 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5041 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5101 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5161 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5221 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5281 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5341 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5401 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5461 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5521 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5581 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5701 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5761 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5821 aaacttggtc ccgggttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5881 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5941 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6001 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6061 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6121 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     6181 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6241 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6301 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6361 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6421 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     6481 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6541 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6601 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6661 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatgttattg
     6721 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6781 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6841 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     6901 agaatt
//
