LOCUS       sequence-393066-        5720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMB1' ori, L-arabinose PBAD promoter, AmpR. Kringle YFP (Yellow 
            Fluorescence Protein) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  AUTHORS   Lozano Terol G, Gallego-Jara J, Sola Martinez RA, Martinez Vivancos 
            A, Canovas Diaz M, de Diego Puente T
  TITLE     Impact of the Expression System on Recombinant Protein Production in
            Escherichia coli BL21.
  JOURNAL   Front Microbiol. 2021 Jun 21;12:682001. doi: 
            10.3389/fmicb.2021.682001. eCollection 2021.
  PUBMED    34234760
REFERENCE   2  (bases 1 to 5720)
  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: "Front 
            Microbiol."; date: "2021-06-21"; volume: "12:682001. doi"; pages: " 
            10.3389/fmicb.2021.682001. eCollection 2021"
COMMENT     Sequence Label: pSF-pMB1'- BAD-YFP
FEATURES             Location/Qualifiers
     source          1..5720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..89
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(94..140)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    158..206
                     /note="synthetic polyadenylation signal"
     polyA_signal    241..289
                     /note="synthetic polyadenylation signal"
     misc_feature    303..394
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     343..362
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             complement(409..1287)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1314..1598
                     /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     1521..1540
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     polyA_signal    2453..2574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2464..2483)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2518..2537
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2689..2735
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2765..2901
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2946..3017
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3243..3831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3323..3342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4577..4645,4646..5437)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4795..4814)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5450..5586
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
       61 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
      121 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
      181 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctgat
      241 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgaaagta
      301 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      361 cccagtgcaa atgcaggtgc cagaacattt ctctgtcgac caactagttt atgacaactt
      421 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
      481 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
      541 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
      601 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
      661 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
      721 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
      781 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
      841 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
      901 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
      961 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     1021 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     1081 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     1141 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     1201 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     1261 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac
     1321 caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct
     1381 aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat
     1441 gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt
     1501 gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag cggatcctac
     1561 ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttt gaattcaaag
     1621 gaggtaccca ccatgacggc actgactgaa ggcgcaaaac tgttcgagaa agaaatccca
     1681 tatatcactg agctggaagg tgacgttgaa ggtatgaagt ttatcatcaa gggtgaaggc
     1741 accggtgacg cgagcgtcgg taaagtggat gctcagttca tttgtaccac gggcgacgtt
     1801 ccggttccgt ggagcacgct ggtcaccacg ctgacgtatg gtgctcagtg ctttgccaag
     1861 tatccgcgcc acattgcgga tttcttcaaa agctgcatgc cggaaggtta cgtccaagag
     1921 cgcaccatca cctttgaggg tgatggcgtg ttcaagaccc gtgcggaagt cacctttgaa
     1981 aatggcagcg tgtacaaccg tgtaaaactg aacggccagg gtttcaagaa ggacggccac
     2041 gtgctgggca aaaatctgga gtttaacttt acccctcatt gtttgtacat ttggggtgac
     2101 caagcgaatc atggcctgaa gagcgcgttc aaaatcatgc atgagatcac cggctccaaa
     2161 gaggatttca ttgttgccga tcacacccaa atgaataccc cgattggtgg tggtccggtg
     2221 cacgtgccgg agtaccacca cattacgtat catgttaccc tgtctaaaga cgtcaccgat
     2281 caccgtgacc atttgaacat tgttgaggtg atcaaggcag ttgacctgga gacgtaccgc
     2341 tagaagttgt ctcctcctgc actgactgac tgatacaatc gatttctgga tccgcaggcc
     2401 tctgctagct tgactgactg agatacagcg taccttcagc tcacagacat gataagatac
     2461 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2521 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2581 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2641 aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc
     2701 ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct
     2761 accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     2821 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     2881 ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat
     2941 caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa
     3001 actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt
     3061 caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc
     3121 agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa
     3181 atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3241 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3301 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3361 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3421 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3481 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3541 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3601 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3661 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3721 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3781 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3841 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3901 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3961 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4021 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg
     4081 aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc
     4141 tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg
     4201 gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag
     4261 agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg
     4321 gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt
     4381 tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata
     4441 ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc
     4501 tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc
     4561 agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4621 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4681 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4741 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     4801 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4861 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     4921 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4981 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5041 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5101 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     5161 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     5221 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5281 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5341 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5401 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     5461 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     5521 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     5581 ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     5641 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     5701 gtcagcttac ctttttggca
//
