LOCUS       sequence-125451-        3202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derived from 21 with silent mutations to make BsaI, BbsI compatible 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3202)
  AUTHORS   Sun ZZ, Yeung E, Hayes CA, Noireaux V, Murray RM
  TITLE     Linear DNA for rapid prototyping of synthetic biological circuits in
            an Escherichia coli based TX-TL cell-free system.
  JOURNAL   ACS Synth Biol. 2014 Jun 20;3(6):387-97. doi: 10.1021/sb400131a. 
            Epub 2013 Dec 4.
  PUBMED    24303785
REFERENCE   2  (bases 1 to 3202)
  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"; doi: "10.1021/sb400131a";
            journalName: "ACS Synth Biol"; date: "2014-06-20- 20"; volume: "3"; 
            issue: "6"; pages: "387-97"
COMMENT     Sequence Label: pBEST_OR2-OR1-Pr_UTR1-deGFP-T500
            BsaI, BbsI-sa...
FEATURES             Location/Qualifiers
     source          1..3202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(66..87)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(327..346)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     674..695
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(890..907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1061..1649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1141..1160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1820..2611,2612..2680))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHATRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2443..2462
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2681..2785)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2853..2871
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatgga gcttttcact ggcgttgttc
       61 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      121 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      181 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
      241 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      301 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      361 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
      421 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
      481 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
      541 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
      601 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
      661 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc taactcgagc
      721 aaagcccgcc gaaaggcggg cttttctgtg tcgaccgatg cccttgagag ccttcaaccc
      781 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtgtt
      841 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
      901 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      961 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1021 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1081 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1141 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1201 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1261 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1321 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1381 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1441 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1501 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1561 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1621 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1681 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1741 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1801 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1861 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1921 agggcttacc atctggcccc agtgctgcaa tgataccgcg tgacccacgc tcaccggctc
     1981 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2041 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2101 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2161 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt gcatgatccc
     2221 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2281 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2341 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2401 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     2461 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2521 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2581 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2641 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     2701 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2761 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     2821 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtg
     2881 ttcaagaatt ctggcgaatc ctctgaccag ccagaaaacg acctttctgt ggtgaaaccg
     2941 gatgctgcaa ttcagagcgc cagcaagtgg gggacagcag atgacctgac cgccgcagag
     3001 tggatgtttg acatggtgat gactatcgca ccatcagcca gaaaaccgaa ttttgctggg
     3061 tgggctaacg atatccgcct gatgcgtgaa cgtgacggac gtaaccaccg cgacatgtgt
     3121 gtgctgttcc gctgggcatg ctgagctaac accgtgcgtg ttgacaattt tacctctggc
     3181 ggtgataatg gttgcagcta gc
//
