LOCUS       sequence-378139-        5956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMB1' ori, lacI, Ptrc promoter, AmpR. Kringle YFP (Yellow 
            Fluorescence Protein) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5956)
  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 5956)
  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'- trc-YFP
FEATURES             Location/Qualifiers
     source          1..5956
                     /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"
     promoter        401..478
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             479..1561
                     /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     complement(498..517)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1793..1798,1799..1815,1816..1822)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1830..1846
                     /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    2689..2810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2700..2719)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2754..2773
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2925..2971
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3001..3137
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3182..3253
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3479..4067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3559..3578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4813..4881,4882..5673)
                     /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(5031..5050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5686..5822
                     /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 gacaccatcg aatggtgcaa
      421 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
      481 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
      541 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
      601 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
      661 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
      721 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
      781 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
      841 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
      901 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
      961 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     1021 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     1081 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     1141 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     1201 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     1261 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
     1321 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     1381 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     1441 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     1501 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     1561 agcgcaacgc aattaatgta agttagcgcg aattgatctg gtttgacagc ttatcatcga
     1621 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     1681 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     1741 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat
     1801 taatcatccg gctcgtataa tgtgtggaat tgtgagcgga taacaagaat tcaaaggagg
     1861 tacccaccat gacggcactg actgaaggcg caaaactgtt cgagaaagaa atcccatata
     1921 tcactgagct ggaaggtgac gttgaaggta tgaagtttat catcaagggt gaaggcaccg
     1981 gtgacgcgag cgtcggtaaa gtggatgctc agttcatttg taccacgggc gacgttccgg
     2041 ttccgtggag cacgctggtc accacgctga cgtatggtgc tcagtgcttt gccaagtatc
     2101 cgcgccacat tgcggatttc ttcaaaagct gcatgccgga aggttacgtc caagagcgca
     2161 ccatcacctt tgagggtgat ggcgtgttca agacccgtgc ggaagtcacc tttgaaaatg
     2221 gcagcgtgta caaccgtgta aaactgaacg gccagggttt caagaaggac ggccacgtgc
     2281 tgggcaaaaa tctggagttt aactttaccc ctcattgttt gtacatttgg ggtgaccaag
     2341 cgaatcatgg cctgaagagc gcgttcaaaa tcatgcatga gatcaccggc tccaaagagg
     2401 atttcattgt tgccgatcac acccaaatga ataccccgat tggtggtggt ccggtgcacg
     2461 tgccggagta ccaccacatt acgtatcatg ttaccctgtc taaagacgtc accgatcacc
     2521 gtgaccattt gaacattgtt gaggtgatca aggcagttga cctggagacg taccgctaga
     2581 agttgtctcc tcctgcactg actgactgat acaatcgatt tctggatccg caggcctctg
     2641 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     2701 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2761 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2821 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2881 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     2941 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     3001 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     3061 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     3121 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     3181 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     3241 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     3301 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     3361 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     3421 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3481 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3541 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3601 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3661 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3721 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3781 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3841 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3901 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3961 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4021 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4081 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4141 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4201 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4261 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     4321 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     4381 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     4441 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     4501 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     4561 gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct
     4621 tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt
     4681 actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat
     4741 cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca
     4801 ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4861 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4921 cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4981 agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc
     5041 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5101 cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca
     5161 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5221 ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5281 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5341 gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta
     5401 cttactctag cttcccggca acagttgata gactggatgg aggcggataa agttgcagga
     5461 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5521 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5581 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5641 gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa
     5701 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     5761 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     5821 aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     5881 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     5941 gcttaccttt ttggca
//
