LOCUS       sequence-48262-e        6011 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 6011)
  AUTHORS   Austin BP, Tozser J, Bagossi P, Tropea JE, Waugh DS
  TITLE     The substrate specificity of Metarhizium anisopliae and Bos taurus 
            carboxypeptidases A: insights into their use as tools for the 
            removal of affinity tags.
  JOURNAL   Protein Expr Purif. 2011 May;77(1):53-61. Epub 2010 Nov 10.
  PUBMED    21073956
REFERENCE   2  (bases 1 to 6011)
  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: "2011-05"; volume: "77(1)"; pages: "53-61. Epub 
            2010 Nov 10"
COMMENT     Sequence Label: p1177-X1-8
FEATURES             Location/Qualifiers
     source          1..6011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..115
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     40..57
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    160..184
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1441..1458
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(1466..1490)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1622..1756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1659..1678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1713..1732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1785..1950
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2133..2588
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2220..2239)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2430..2451
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2615..2719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2720..2788,2789..3580)
                     /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(2938..2957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3751..4339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4240..4259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4493..4510
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4642..4866
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4933..5466)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5655..5660,5661..5677,5678..5683))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 cgtatactcc ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa
       61 taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc
      121 ggattattca taccgtccca ccatcgggcg cggatcatca caagtttgta caaaaaagtt
      181 ggcaccatga gagtggttgc tttcttcgcc tgtctagtag ccacggcggc cacaatgccg
      241 gctgagagcc ctgtgtctta cgacgggtac aaggtctttc gagtcccggt cgtggacgat
      301 ggcacgcaca tccagagtct tattgatcat ttgaatctga atgtctggca gccgccctcc
      361 aaaaagggtg cttttgccga catccaggtg gccccaagcc agttggcagc cttcgaaaac
      421 gcaatgaaag ggcgcagctt cgagattatg catgaggatc tcggagactc cattgccaga
      481 gaaggcacca tccaggcata tgcggccggc tcggctaatg cctcctggtt cacatcgtac
      541 catccataca atgaccatct tcaatggatg aaggacattg cctctcaata cccgagcaat
      601 gttaagagtg tcacgtctgg aaccacgggt gatggcaata ccataacggg cctgcatatt
      661 ttcggcagca gcggtggcgg caataaaccc gccgtcgttt ttcatggcac cgtccatgcg
      721 cgcgaatgga tcgttgctat gactctcgag tatataacca acgagctcct cgccaagtac
      781 gctaccgaca gcgccgtaaa ggctgttgtg gacaagtacg acttctacat gttccccatt
      841 gtcaatgttg acggcttcaa gtacacgcag tcgtcagacc gcatgtggag gaagaaccgg
      901 tctcgcaacc agggctcgag ctgtctcggc acggatccca atcgcaactg gccatacaag
      961 tgggatggcc ctgggtcatc aacgaaccct tgtactgaga cgtatcgggg tgcatctgcg
     1021 ggcaacagcc cagaagtcaa gtcgtacatc gccttcctcg acaagatcaa gaagagccaa
     1081 ggcgtcaagc tatacatcga ctggcactcg tattctcagc tgttcatgac accatacggg
     1141 tactcgtgca gcgcaaggac gcccaacaat gctgccttgc aagctctggc aaagggtgct
     1201 tctgacgcta tgaggagtgt ccacggaacc accttcgcat atggtccagt ttgcaatgtc
     1261 atctaccaag ttgccggtgg ctctattgat tgggtccaag acgttcttaa agccgacaat
     1321 gtcttcacaa tcgaactgcg tgacaaaggt aggtatgggt ttgtcctgcc accggaccag
     1381 atcataccgt ctggagagga gagctttgct ggtgccatgc atctcttcca gcagatgagt
     1441 caccatcacc atcaccatcg tcgctagcca actttcttgt acaaagtggt gatagcttgt
     1501 cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt
     1561 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1621 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1681 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1741 ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag ataagtgaaa
     1801 tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca
     1861 gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct
     1921 cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt
     1981 aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact ttttctctgt
     2041 cacagaatga aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat
     2101 caacgcttat ttgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     2161 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2221 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2281 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2341 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2401 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2461 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2521 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     2581 tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2641 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2701 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2761 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2821 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2881 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2941 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3001 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3061 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3121 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3181 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3241 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3301 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3361 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3421 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3481 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3541 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3601 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3661 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3721 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3781 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3841 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3901 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3961 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4021 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4081 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4141 gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4201 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4261 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4321 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4381 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4441 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4501 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4561 tatttcacac cgcagaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
     4621 tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga
     4681 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
     4741 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
     4801 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat ggtaaagact
     4861 atattcgcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc
     4921 ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg
     4981 tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta
     5041 gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc
     5101 aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg
     5161 cggctgctca aacctgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc
     5221 gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc
     5281 cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc
     5341 agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact
     5401 tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg
     5461 taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct
     5521 tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca
     5581 ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac
     5641 gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca
     5701 tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gaggcatttc
     5761 tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag gcattggcgg
     5821 ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt caggagatcg
     5881 gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa gtggttcgca
     5941 tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc tatagttcta
     6001 gtggttggct a
//
