LOCUS       sequence-382466-        5729 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5729)
  TITLE     8-series E. coli arabinose expression vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5729)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "Unpublished"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 Strep His6 TEV LIC cloning vector (8HR)
FEATURES             Location/Qualifiers
     source          1..5729
                     /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(298..318)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(325..348)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(364..381)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             401..423
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(457..741)
                     /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(515..534)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             768..1646
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(1805..1824)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3282..3473
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3467..3489)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3575..3715
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3630..3649
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3730..3747)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3901..4489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3981..4000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4659..5450,5451..5519))
                     /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     5282..5301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5520..5624)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5692..5710
                     /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 aggttttcgg cgcctttttc gaactgcggg tggctccagc tagcagaaga
      361 accatggtga tggtgatggt gagaagattt catatgtata tctccttctt aaagttaaac
      421 aaaattattt ctagatgtta gcccaaaaaa acgggtatgg agaaacagta gagagttgcg
      481 ataaaaagcg tcaggtaaga tccgctaatc ttatggataa aaatgctatg gcatagcaaa
      541 gtgtgacgcc gtgcaaataa tcaatgtgga cttttctgcc gtgattatag acacttttgt
      601 tacgcgtttt tgtcatggct ttggtcccgc tttgttacag aatgctttta ataagcgggg
      661 ttaccggttt ggttagcgag aagagccagt aaaagacgca gtgacggcaa tgtctgatgc
      721 aatatggaca attggtttct tctctgaatg gcgggagtat gaaaagtatg gctgaagcgc
      781 aaaatgatcc cctgctgccg ggatactcgt ttaatgccca tctggtggcg ggtttaacgc
      841 cgattgaggc caacggttat ctcgattttt ttatcgaccg accgctggga atgaaaggtt
      901 atattctcaa tctcaccatt cgcggtcagg gggtggtgaa aaatcaggga cgagaatttg
      961 tttgccgacc gggtgatatt ttgctgttcc cgccaggaga gattcatcac tacggtcgtc
     1021 atccggaggc tcgcgaatgg tatcaccagt gggtttactt tcgtccgcgc gcctactggc
     1081 atgaatggct taactggccg tcaatctttg ccaatacggg gttctttcgc ccggatgaag
     1141 cgcaccagcc gcatttcagc gacctgtttg ggcaaatcat taacgccggg caaggggaag
     1201 ggcgctattc ggagctgctg gcgataaatc tgcttgagca attgttgctg cggcgcatgg
     1261 aagcgattaa cgagtcgctc catccaccga tggataatcg ggtacgcgag gcttgtcagt
     1321 acatcagcga tcacctggca gacagcaatt ttgatatagc cagcgtcgca cagcatgttt
     1381 gcttgtcgcc gtcgcgtctg tcacgtcttt tccgccagca gttagggatt agcgtcttaa
     1441 gctggcgcga ggaccaacgt atcagccagg cgaagctgct tttgagcacc acccggatgc
     1501 ctatcgccac cgtcggtcgc aatgttggtt ttgacgatca actctatttc tcgcgggtat
     1561 ttaaaaaatg caccggggcc agcccgagcg agttccgtgc cggttgtgaa gaaaaagtga
     1621 atgatgtagc cgtcaagttg tcataattgg taacgaatca gacaattgac ggctcgaggg
     1681 agtagcatag ggtttgcaga atccctgctt cgtccatttg acgggcacat tatggcggcc
     1741 gcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     1801 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     1861 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
     1921 atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
     1981 ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gaccgatgcc cttgagagcc
     2041 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     2101 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     2161 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     2221 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     2281 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     2341 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     2401 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     2461 caaggatcgc tcgcggctct taccagccta acttcgatca ttggaccgct gatcgtcacg
     2521 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc
     2581 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc
     2641 tgaatggaag ccggcggcac ctcgctaacg gattcaccac tccaagaatt ggagccaatc
     2701 aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt
     2761 ccgccatctc cagcagccgc acgcggcgca tctcgggcag cgttgggtcc tggccacggg
     2821 tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg
     2881 gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt
     2941 ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa
     3001 cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc
     3061 taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt
     3121 ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc
     3181 gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca
     3241 ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa
     3301 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca
     3361 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg
     3421 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3481 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3541 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     3601 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     3661 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     3721 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3781 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3841 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4081 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4321 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4501 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4561 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4621 atcaatctaa agtatatatg agtaaacttg gtcccgggtt accaatgctt aatcagtgag
     4681 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4741 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4801 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4861 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4921 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     4981 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5041 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5101 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5161 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5221 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     5281 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5341 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5401 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5461 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5521 ctcttccttt ttcaatgtta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5581 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5641 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5701 atcacgaggc cctttcgtct tcaagaatt
//
