LOCUS       sequence-378192-        5526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  p15A ori, lacI, Ptac promoter, AmpR. Kringle YFP (Yellow 
            Fluorescence Protein) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5526)
  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 5526)
  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-p15A-tac- YFP
FEATURES             Location/Qualifiers
     source          1..5526
                     /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(1678..1683,1684..1699,1700..1706)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1714..1730
                     /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)."
     primer_bind     1719..1741
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     polyA_signal    2615..2736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2626..2645)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2680..2699
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2851..2897
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2927..3063
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3108..3179
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3274..3819)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3354..3373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4383..4451,4452..5243)
                     /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(4601..4620)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5256..5392
                     /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 actgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     1621 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
     1681 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
     1741 gaaacagaat tcccggaagc ttccgagctc tcgaattcaa aggaggtacc caccatgacg
     1801 gcactgactg aaggcgcaaa actgttcgag aaagaaatcc catatatcac tgagctggaa
     1861 ggtgacgttg aaggtatgaa gtttatcatc aagggtgaag gcaccggtga cgcgagcgtc
     1921 ggtaaagtgg atgctcagtt catttgtacc acgggcgacg ttccggttcc gtggagcacg
     1981 ctggtcacca cgctgacgta tggtgctcag tgctttgcca agtatccgcg ccacattgcg
     2041 gatttcttca aaagctgcat gccggaaggt tacgtccaag agcgcaccat cacctttgag
     2101 ggtgatggcg tgttcaagac ccgtgcggaa gtcacctttg aaaatggcag cgtgtacaac
     2161 cgtgtaaaac tgaacggcca gggtttcaag aaggacggcc acgtgctggg caaaaatctg
     2221 gagtttaact ttacccctca ttgtttgtac atttggggtg accaagcgaa tcatggcctg
     2281 aagagcgcgt tcaaaatcat gcatgagatc accggctcca aagaggattt cattgttgcc
     2341 gatcacaccc aaatgaatac cccgattggt ggtggtccgg tgcacgtgcc ggagtaccac
     2401 cacattacgt atcatgttac cctgtctaaa gacgtcaccg atcaccgtga ccatttgaac
     2461 attgttgagg tgatcaaggc agttgacctg gagacgtacc gctagaagtt gtctcctcct
     2521 gcactgactg actgatacaa tcgatttctg gatccgcagg cctctgctag cttgactgac
     2581 tgagatacag cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa
     2641 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2701 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2761 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2821 tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg
     2881 tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa
     2941 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     3001 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     3061 taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg
     3121 gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa
     3181 tctcattggt taccttgggc tatcgaaact taattaagga agatgccagg aagatactta
     3241 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     3301 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     3361 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     3421 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     3481 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
     3541 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
     3601 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     3661 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     3721 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     3781 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaagggg
     3841 tctgacgctc agtggaacga aaaatttggt ggtatttaaa tttccgaact ctccaaggcc
     3901 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     3961 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     4021 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     4081 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     4141 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     4201 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     4261 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     4321 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     4381 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4441 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4501 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4561 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4621 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4681 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4741 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4801 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4861 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4921 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     4981 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     5041 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5101 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5161 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5221 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     5281 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     5341 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     5401 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     5461 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     5521 ttggca
//
