LOCUS       sequence-56137-e        2966 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 2966)
  AUTHORS   Korendovych IV, Kulp DW, Wu Y, Cheng H, Roder H, DeGrado WF
  TITLE     Design of a switchable eliminase.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Apr 26;108(17):6823-7. Epub 2011 Apr 
            11.
  PUBMED    21482808
REFERENCE   2  (bases 1 to 2966)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2011-04-26"; volume: "108(17)"; pages: 
            "6823-7. Epub 2011 Apr 11"
COMMENT     Sequence Label: pEXP5-NT-AlleyCat
FEATURES             Location/Qualifiers
     source          1..2966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             122..142
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(422..440)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      436..483
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(528..546)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        614..718
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(719..787,788..1579)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(937..956)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1750..2338
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2239..2258
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2492..2509
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2589..2608)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacca tgtctggttc tcatcatcat catcatcatg gtagcagcgg
      121 cgaaaacctg tattttcagt cccttatgaa agatacagat agcgaagaag aaattagaga
      181 agcgttccgt gtggaagaca aggatggtaa tggttacatt agtgctgcag aacttcgtca
      241 tgtgatgaca aatcttgggg agaagctaac agatgaagaa gttgatgaaa tgattaggga
      301 agcagacatt gatggtgatg gtcaagtaaa ctatgaagag tttgtacaaa tgatgacagc
      361 gaagtgaagg gtgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
      421 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      481 ttgctgaaag gaggaactat atccggataa ttcttgaaga cgaaagggcc tcgtgatacg
      541 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
      601 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      661 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      721 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      781 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      841 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      901 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      961 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1021 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1081 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1141 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1201 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1261 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1321 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1381 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1441 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1501 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1561 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1621 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1681 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1741 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1801 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1861 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1921 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1981 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2041 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2101 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2161 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2221 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2281 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2341 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2401 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2461 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2521 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     2581 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     2641 tacgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     2701 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     2761 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     2821 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     2881 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     2941 tgcccgagat ctcgatcccg cgaaat
//
