LOCUS       Exported                3089 bp DNA     circular SYN 23-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBSM
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3089)
  AUTHORS   Ponchon L, Catala M, Seijo B, El Khouri M, Dardel F, Nonin-Lecomte 
            S, Tisne C
  TITLE     Co-expression of RNA-protein complexes in Escherichia coli and 
            applications to RNA biology.
  JOURNAL   Nucleic Acids Res. 2013 Aug;41(15):e150. doi: 10.1093/nar/gkt576. 
            Epub 2013 Jun 26.
  PUBMED    23804766
REFERENCE   2  (bases 1 to 3089)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 23, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2013-08"; volume: "41(15):e150. doi"; pages: " 
            10.1093/nar/gkt576. Epub 2013 Jun 26"
FEATURES             Location/Qualifiers
     source          1..3089
                     /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"
     promoter        complement(279..297)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(318..334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(318..334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(331..353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    342..358
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(366..396)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    411..432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(549..566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(720..1308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(800..819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1479..2339)
                     /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     2102..2121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2340..2444)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2470..2925)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2607..2628)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2819..2838
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3052..3074
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3066..3083
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3067..3083
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattggag ctaaagcttg gagctccgga tcctaagccc
       61 ccatcaagct taaaaaaaat ccttagcttt cgctaaggat ctgcagtggt ggctacgacg
      121 ggattcgaac ctgtgacccg cggatatcgg ccgctgctct aaccaactga gctacgtagc
      181 cgaattcagc gttacaagta ttacacaaag ttttttatgt tgagaatatt tttttgatgg
      241 ggcgacctcg agggggggcc cggtaccagc ttttgttccc tttagtgagg gttaatttcg
      301 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
      361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
      421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
      481 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
      541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1141 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1381 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     1441 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     1501 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     1561 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     1621 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1681 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1741 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     1801 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     1861 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1921 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1981 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2041 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     2101 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     2161 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     2221 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     2281 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     2341 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     2401 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     2461 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2521 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2581 agaccgagat agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2641 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2701 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2761 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2821 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2881 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     2941 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3001 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     3061 gacgttgtaa aacgacggcc agtgaattg
//
