LOCUS       sequence-378141-        4257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  p15A ori, promotorless, AmpR. Kringle YFP (Yellow Fluorescence 
            Protein) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4257)
  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 4257)
  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-YFP
FEATURES             Location/Qualifiers
     source          1..4257
                     /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"
     polyA_signal    1346..1467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1357..1376)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1411..1430
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1582..1628
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1658..1794
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1839..1910
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2005..2550)
                     /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(2085..2104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3114..3182,3183..3974)
                     /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(3332..3351)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3987..4123
                     /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 caactagtac cccgggtagg
      421 gcccaattcg aagtagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
      481 cgctgccaag cttccgagct ctcgaattca aaggaggtac ccaccatgac ggcactgact
      541 gaaggcgcaa aactgttcga gaaagaaatc ccatatatca ctgagctgga aggtgacgtt
      601 gaaggtatga agtttatcat caagggtgaa ggcaccggtg acgcgagcgt cggtaaagtg
      661 gatgctcagt tcatttgtac cacgggcgac gttccggttc cgtggagcac gctggtcacc
      721 acgctgacgt atggtgctca gtgctttgcc aagtatccgc gccacattgc ggatttcttc
      781 aaaagctgca tgccggaagg ttacgtccaa gagcgcacca tcacctttga gggtgatggc
      841 gtgttcaaga cccgtgcgga agtcaccttt gaaaatggca gcgtgtacaa ccgtgtaaaa
      901 ctgaacggcc agggtttcaa gaaggacggc cacgtgctgg gcaaaaatct ggagtttaac
      961 tttacccctc attgtttgta catttggggt gaccaagcga atcatggcct gaagagcgcg
     1021 ttcaaaatca tgcatgagat caccggctcc aaagaggatt tcattgttgc cgatcacacc
     1081 caaatgaata ccccgattgg tggtggtccg gtgcacgtgc cggagtacca ccacattacg
     1141 tatcatgtta ccctgtctaa agacgtcacc gatcaccgtg accatttgaa cattgttgag
     1201 gtgatcaagg cagttgacct ggagacgtac cgctagaagt tgtctcctcc tgcactgact
     1261 gactgataca atcgatttct ggatccgcag gcctctgcta gcttgactga ctgagataca
     1321 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
     1381 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1441 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1501 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     1561 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     1621 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     1681 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     1741 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     1801 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     1861 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     1921 ttaccttggg ctatcgaaac ttaattaagg aagatgccag gaagatactt aacagggaag
     1981 tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga
     2041 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2101 tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat
     2161 tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag
     2221 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat
     2281 ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag
     2341 ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact
     2401 gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg
     2461 gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa
     2521 gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct
     2581 cagtggaacg aaaaatttgg tggtatttaa atttccgaac tctccaaggc cctcgtcgga
     2641 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2701 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2761 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2821 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     2881 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     2941 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     3001 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     3061 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     3121 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3181 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     3241 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3301 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3361 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3421 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3481 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     3541 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3601 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3661 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     3721 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3781 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3841 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3901 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3961 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     4021 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     4081 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     4141 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     4201 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggca
//
