LOCUS       sequence-48267-e        3619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3619)
  AUTHORS   Austin BP, Waugh DS
  TITLE     Isolation of Metarhizium anisopliae carboxypeptidase A with native 
            disulfide bonds from the cytosol of Escherichia coli BL21(DE3).
  JOURNAL   Protein Expr Purif. 2012 Mar;82(1):116-24. Epub 2011 Dec 14.
  PUBMED    22197595
REFERENCE   2  (bases 1 to 3619)
  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: "Protein 
            Expr Purif."; date: "2012-03"; volume: "82(1)"; pages: "116-24. Epub
            2011 Dec 14"
COMMENT     Sequence Label: pBA2219
FEATURES             Location/Qualifiers
     source          1..3619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      108..653
                     /direction=RIGHT
                     /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     554..573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     770..787
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1237..1256)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1248..2063
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1248..2063
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1321..1340)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2665..2684
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2665..2683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2715..2737
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2744..2776
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     protein_bind    2786..2810
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3460..3484)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3533..3551)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3547..3594
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
       61 gagttttcgt tccactgagc gtcagacccc ttaataagat gatcttcttg agatcgtttt
      121 ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg
      181 aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca
      241 ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct
      301 aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc
      361 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca
      421 gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac
      481 gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga
      541 gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
      601 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
      661 ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc
      721 gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt
      781 gagcgaggaa gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt
      841 ttttctcctg ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc
      901 agtatacact ccgctagcgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc
      961 aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt
     1021 tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt
     1081 tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa
     1141 gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     1201 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     1261 aacgggaaac gtcttgctcg aggccgcgat taaattccaa catggatgct gatttatatg
     1321 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
     1381 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     1441 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     1501 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa
     1561 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     1621 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     1681 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     1741 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc
     1801 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     1861 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     1921 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     1981 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     2041 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     2101 ttacgctgac ttgacgggac ggcggctttg ttgaataaat cgaacttttg ctgagttgaa
     2161 ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa
     2221 aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg
     2281 gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag
     2341 acctcagcgc tcaaagatgc aggggtaaaa gctaaccgca tctttaccga caaggcatcc
     2401 ggcagttcaa cagatcggga agggctggat ttgctgagga tgaaggtgga ggaaggtgat
     2461 gtcattctgg tgaagaagct cgaccgtctt ggccgcgaca ccgccgacat gatccaactg
     2521 ataaaagagt ttgatgctca gggtgtagcg gttcggttta ttgacgacgg gatcagtacc
     2581 gacggtgata tggggcaaat ggtggtcacc atcctgtcgg ctgtggcaca ggctgaacgc
     2641 cggaggatct cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc
     2701 cctctagaaa taattttgtt taactttaag aaggagatat accatggcta gcatgactgg
     2761 tggacagcaa atgggtcgga tcatcacaag tttgtacaaa aaagttgtgg acgacgcggc
     2821 aattcaacaa acgttagcca aaatgggcat caaaagcagc gatattcagc ccgcgcctgt
     2881 agctggcatg aagacagttc tgactaacag cggcgtgttg tacatcaccg atgatggtaa
     2941 acatatcatt caggggccaa tgtatgacgt tagtggcacg gctccggtca atgtcaccaa
     3001 taagatgctg ttaaagcagt tgaatgcgct tgaaaaagag atgatcgttt ataaagcgcc
     3061 gcaggaaaaa cacgtcatca ccgtgtttac tgatattacc tgtggttact gccacaaact
     3121 gcatgagcaa atggcagact acaacgcgct ggggatcacc gtgcgttatc ttgctttccc
     3181 gcgccagggg ctggacagcg atgcagagaa agaaatgaaa gctatctggt gtgcgaaaga
     3241 taaaaacaaa gcgtttgatg atgtgatggc aggtaaaagc gtcgcaccag ccagttgcga
     3301 cgtggatatt gccgaccatt acgcacttgg cgtccagctt ggcgttagcg gtactccggc
     3361 agttgtgctg agcaatggca cacttgttcc gggttaccag ccgccgaaag agatgaaaga
     3421 attcctcgac gaacaccaaa aaatgaccag cggtaaataa tgcaactttc ttgtacaaag
     3481 tggtgatcgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     3541 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     3601 ggaggaacta tatccggat
//
