LOCUS       sequence-36713-e        2950 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 2950)
  AUTHORS   Pereira MJ, Behera V, Walter NG
  TITLE     Nondenaturing purification of co-transcriptionally folded RNA avoids
            common folding heterogeneity.
  JOURNAL   PLoS One. 2010  . 5(9):e12953.
  PUBMED    20886091
REFERENCE   2  (bases 1 to 2950)
  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: "PLoS One. 
            2010  . 5(9):e12953."
COMMENT     Sequence Label: pMCGL
FEATURES             Location/Qualifiers
     source          1..2950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(65..84)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(66..84)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(103..119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(103..119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(116..138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    127..143
                     /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(join(151..157,158..175,176..181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    196..217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(334..351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(505..1093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(585..604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1264..2052)
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     1887..1906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2124..2228)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2296..2314
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2352..2374)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2474..2493
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2687..2709
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2701..2718
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2702..2718
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2726..2744
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2726..2743
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttcgggcgcg catgcctgca ggtcgactct agaggatccc cgggtaccga gctcaagctt
       61 gtctccctat agtgagtcgt attagagctt ggcgtaatca tggtcatagc tgtttcctgt
      121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
      301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
      361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
      421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
      961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     1441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     1921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     1981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2041 cagcgtttct gggtgagcaa aaacaggaag gcaaatgccg caaaaaaggg aataagggcg
     2101 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2161 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2221 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     2281 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     2341 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     2401 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     2461 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     2521 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     2581 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     2641 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     2701 tgtaaaacga cggccagtga attggattta ggtgacacta tagaatacga attccgcagg
     2761 cctgctcgga tcatgtgatt tctctttgtt caaggagtca cccccttggt ttgaagaaat
     2821 ccttacggct gtgatctgca cactcagccg ggaggcatcc gccgaaaatt cgataacctc
     2881 catcctcgtc aacttaaacc gcggggcgcg actatcttac gcacttgcat gattctcctc
     2941 gctgcttaag
//
