LOCUS       sequence-333644-        2683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TrrnB and L3S2P21 terminator part (D).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2683)
  AUTHORS   Greco FV, Pandi A, Erb TJ, Grierson CS, Gorochowski TE
  TITLE     Harnessing the central dogma for stringent multi-level control of 
            gene expression.
  JOURNAL   Nat Commun. 2021 Mar 19;12(1):1738. doi: 10.1038/s41467-021-21995-7.
  PUBMED    33741937
REFERENCE   2  (bases 1 to 2683)
  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: "Nat 
            Commun."; date: "2021-03-19"; volume: "12(1):1738. doi"; pages: " 
            10.1038/s41467-021-21995-7"
COMMENT     Sequence Label: pD-01
FEATURES             Location/Qualifiers
     source          1..2683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..67)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(160..748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(240..259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(919..1710,1711..1779))
                     /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     1542..1561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1780..1884)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2185..2207
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2199..2216
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2200..2216
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2227..2246
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2227..2245
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2298..2384
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2476..2503
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 attaacatgg tcatagctgt ttccttgcgt attgggcgct ctccgcttcc tcgctcactg
       61 actcgctgcg ctcggtcgtt cgggtaaagc ctggggtgcc taatgagcaa aaggccagca
      121 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      181 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      241 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      301 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      361 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      421 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      481 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      541 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      601 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      661 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      721 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      781 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      841 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      901 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      961 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1021 gggcttacca tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc
     1081 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1141 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1201 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1261 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1321 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1381 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1441 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     1501 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     1561 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     1621 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     1681 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     1741 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     1801 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     1861 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     1921 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     1981 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2041 ggccgctaca gggcgctccc attcgccatt caggctgcgc aactgttggg aagggcgttt
     2101 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2161 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     2221 gcgacgtaat acgactcact atagggcgaa ttggcggaag gccgtcaagg ccgcatgaag
     2281 ggcgcgccag gtctcagctt ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     2341 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     2401 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     2461 ggcatcaaat taagcagaag gccatcctga cggatggcct tttggccccc gacccttaga
     2521 ctctgtactc agggaccaaa acgaaaaaag gccccccttt cgggaggcct cttttctgga
     2581 atttggtacc gagcgcttga gaccttttaa ttaacaacct gggcctcatg ggccttccgc
     2641 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgc
//
