LOCUS       sequence-69762-e        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  JOURNAL   Unpublished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            "Unpublished"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"
     primer_bind     complement(67..86)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      879..1467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1368..1387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1621..1638
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1653..1793
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1719..1738)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1879..1901
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1895..2086)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3544..3563
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3722..4600)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4627..4911
                     /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     4834..4853
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4974..4996
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5016..5033
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5043..6143
                     /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     6117..6140
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             6198..6218
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(6308..6325)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6478..6524
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6564..6582)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6650..6754
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6755..6823,6824..6906,1..709)
                     /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"
ORIGIN
        1 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
       61 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      121 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      181 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      241 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      301 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      361 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      421 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      481 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      541 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      601 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
      661 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac ccgggaccaa
      721 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
      781 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
      841 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
      901 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      961 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1021 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1081 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1141 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1201 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1261 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1321 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1381 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1441 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1501 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     1561 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1621 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     1681 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     1741 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     1801 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     1861 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     1921 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     1981 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     2041 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     2101 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     2161 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     2221 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     2281 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     2341 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     2401 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     2461 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     2521 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc
     2581 gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt
     2641 gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt
     2701 tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg
     2761 caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca
     2821 tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccagtga tcgaagttag
     2881 gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct
     2941 ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat
     3001 ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc
     3061 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     3121 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     3181 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     3241 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     3301 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac gctctccctt
     3361 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     3421 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     3481 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     3541 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     3601 cacgatgcgt ccggcgtaga ggatcgcgac cgccataatg tgcctgtcaa atggacgaag
     3661 cagggattct gcaaacccta tgctactccc tcgagccgtc aattgtctga ttcgttacca
     3721 attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc
     3781 tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac
     3841 caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct
     3901 ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa
     3961 gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggct atatcaaaat
     4021 tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg
     4081 gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat
     4141 ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa
     4201 acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa
     4261 agattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact
     4321 ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga
     4381 acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct
     4441 gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa
     4501 aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg
     4561 agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat
     4621 tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact
     4681 ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg
     4741 ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc
     4801 acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat
     4861 tagcggatct tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt
     4921 tttgggctat taattaagcg gccgccctgc aggactcgag ttctagaaat aattttgttt
     4981 aactttaaga aggagatata catatgaaat cttctcacca tcaccatcac catggttctt
     5041 ctatgaaaat cgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg
     5101 gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc
     5161 atccggataa actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca
     5221 ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa
     5281 tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt
     5341 acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca
     5401 aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac
     5461 tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc
     5521 cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta
     5581 aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga
     5641 ttaaaaacaa acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata
     5701 aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca
     5761 aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg
     5821 ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt
     5881 tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc
     5941 tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg
     6001 ccactatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt
     6061 tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg
     6121 aagccctgaa agacgcgcag actaattcga gctcgaacaa caacaacaat aacaataaca
     6181 acaacctcgg gatcgaggaa aacctgtact tccaatccaa tattggaagt ggataacgga
     6241 tccgcgatcg cggcgcgcca cctggtggcc ggcctggctg ttttggcgga tgagagaaga
     6301 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     6361 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     6421 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     6481 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     6541 aactaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     6601 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6661 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6721 ataaatgctt caataacatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6781 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6841 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6901 caacag
//
