LOCUS       sequence-378138-        6044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMB1' ori, lacI PT7 promoter, AmpR. Kringle YFP (Yellow Fluorescence
            Protein) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6044)
  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 6044)
  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'-t7- YFP
FEATURES             Location/Qualifiers
     source          1..6044
                     /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(401..1483)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1445..1464
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1484..1561)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1767..1786
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1870..1889
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1870..1888
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1889..1913
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     polyA_signal    2777..2898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2788..2807)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2842..2861
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3013..3059
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3089..3225
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3270..3341
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3567..4155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3647..3666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4901..4969,4970..5761)
                     /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(5119..5138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5774..5910
                     /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 tcactgcccg ctttccagtc
      421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      481 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
      541 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
      601 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
      661 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
      721 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
      781 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
      841 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
      901 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
      961 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     1021 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     1081 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     1141 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     1201 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     1261 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     1321 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     1381 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     1441 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     1501 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     1561 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     1621 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     1681 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     1741 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     1801 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     1861 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctaa
     1921 gcttccgagc tctcgaattc aaaggaggta cccaccatga cggcactgac tgaaggcgca
     1981 aaactgttcg agaaagaaat cccatatatc actgagctgg aaggtgacgt tgaaggtatg
     2041 aagtttatca tcaagggtga aggcaccggt gacgcgagcg tcggtaaagt ggatgctcag
     2101 ttcatttgta ccacgggcga cgttccggtt ccgtggagca cgctggtcac cacgctgacg
     2161 tatggtgctc agtgctttgc caagtatccg cgccacattg cggatttctt caaaagctgc
     2221 atgccggaag gttacgtcca agagcgcacc atcacctttg agggtgatgg cgtgttcaag
     2281 acccgtgcgg aagtcacctt tgaaaatggc agcgtgtaca accgtgtaaa actgaacggc
     2341 cagggtttca agaaggacgg ccacgtgctg ggcaaaaatc tggagtttaa ctttacccct
     2401 cattgtttgt acatttgggg tgaccaagcg aatcatggcc tgaagagcgc gttcaaaatc
     2461 atgcatgaga tcaccggctc caaagaggat ttcattgttg ccgatcacac ccaaatgaat
     2521 accccgattg gtggtggtcc ggtgcacgtg ccggagtacc accacattac gtatcatgtt
     2581 accctgtcta aagacgtcac cgatcaccgt gaccatttga acattgttga ggtgatcaag
     2641 gcagttgacc tggagacgta ccgctagaag ttgtctcctc ctgcactgac tgactgatac
     2701 aatcgatttc tggatccgca ggcctctgct agcttgactg actgagatac agcgtacctt
     2761 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2821 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2881 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2941 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     3001 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     3061 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     3121 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     3181 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     3241 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     3301 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     3361 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     3421 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     3481 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     3541 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3601 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3661 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3721 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3781 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3841 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3901 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3961 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4021 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4081 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4141 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4201 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4261 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4321 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4381 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     4441 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     4501 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     4561 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     4621 gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga tcctctcagt
     4681 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     4741 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     4801 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     4861 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     4921 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4981 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     5041 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     5101 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5161 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     5221 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5281 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     5341 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5401 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5461 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     5521 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5581 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5641 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5701 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5761 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     5821 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     5881 gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc
     5941 tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc
     6001 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggca
//
