LOCUS       sequence-51565-e        3526 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 3526)
  AUTHORS   Muona M, Aranko AS, Iwai H
  TITLE     Segmental isotopic labelling of a multidomain protein by protein 
            ligation by protein trans-splicing.
  JOURNAL   Chembiochem. 2008 Dec 15. 9(18):2958-61.
  PUBMED    19031436
REFERENCE   2  (bases 1 to 3526)
  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: 
            "Chembiochem. 2008 Dec 15. 9(18):2958-61."
COMMENT     Sequence Label: pHYRS52
FEATURES             Location/Qualifiers
     source          1..3526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(728..746)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      742..789
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      886..1341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(973..992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1183..1204
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1422..1526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1527..1595,1596..2387)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRPPRYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2558..3146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3047..3066
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3300..3317
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3401..3420
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3401..3419
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3451..3473
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3493..3510
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(3514..3526,1..5)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 gatcccttgt tcctgaatta aatgaaaaag acgatgacca agtacaaaaa gctttggcat
       61 ctagagaaaa tactcagtta atgaatagag ataatataga gataacagta cgtgatttta
      121 agaccttggc accacgaaga tggctaaatg acactatcat tgagtttttt atgaaataca
      181 ttgaaaaatc tacccctaat acagtggcgt ttaattcgtt tttctatacc aatttatcag
      241 aaaggggtta tcaaggcgtc cggaggtgga tgaagagaaa gaagacacaa attgataaac
      301 ttgataaaat ctttacacca ataaatttga accaatccca ctgggcgttg ggcataattg
      361 atttaaaaaa gaaaactata ggttacgtag attcattatc gaatggtcca aatgctatga
      421 gtttcgctat actgactgac ttgcaaaaat atgttatgga ggaaagtaag catacaatag
      481 gagaagactt tgatttgatt catttagatt gtccgcagca accaaatggc tacgactgtg
      541 gaatatatgt ttgtatgaat actctctatg gaagtgcaga tgcgccattg gattttgatt
      601 ataaagatgc gattaggatg agaagattta ttgcccattt gattttaacc gacgctttaa
      661 aataagaatt cgaagcttga tccggctgct aacaaagccc gaaaggaagc tgagttggct
      721 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
      781 ggttttttgc tgaaaggagg aactatatcc ggatctggcg taatagcgaa gaggcccgca
      841 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
      901 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      961 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1021 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagagct ttacggcacc
     1081 tcgaccgcaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     1141 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1201 ctggaacaac actcaaccct atcgcggtct attcttttga tttataaggg attttgccga
     1261 tttcggccta ttggttaaaa aatgagctga tttaacaaat atttaacgcg aattttaaca
     1321 aaatattaac gtttacaatt tcgcctgatg cggtattttc tccttacgca tctgtgcggt
     1381 atttcacacc gcatacaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1441 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1501 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1561 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1621 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1681 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1741 ttctgctatg tgatacacta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1801 ccccgcggta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1861 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1921 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1981 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2041 caaacgacga gagtgacacc acgatgcctg tagcaatgcc aacaacgttg cgcaaactat
     2101 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2161 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2221 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2281 agcgctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2341 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2401 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2461 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2521 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2581 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2641 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2701 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2761 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2821 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2881 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2941 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3001 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3061 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3121 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3181 gcttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3241 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3301 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3361 gttggccgat tcattaatgc agatctcgat cccgcgaaat taatacgact cactataggg
     3421 agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg agatatacat
     3481 atgcggggtt ctcatcatca tcatcatcat ggtctggttc cgcgtg
//
