LOCUS       sequence-373300-        2806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector for generating part plasmids, with mNeonGreen dropout
            following BsmBI digestion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 799 to 833)
  AUTHORS   Chen R, Wang SK, Belk JA, Amaya L, Li Z, Cardenas A, Abe BT, Chen 
            CK, Wender PA, Chang HY
  TITLE     Engineering circular RNA for enhanced protein production.
  JOURNAL   Nat Biotechnol. 2022 Jul 18. pii: 10.1038/s41587-022-01393-0. doi: 
            10.1038/s41587-022-01393-0.
  PUBMED    35851375
REFERENCE   2  (bases 1 to 2806)
  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 
            Biotechnol. 2022 Jul 18. pii: 10.1038/s41587-022-01393-0. doi: 
            10.1038/s41587-022-01393-0."
COMMENT     Sequence Label: pRC0569 (parts entry vector)
FEATURES             Location/Qualifiers
     source          1..2806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      92..680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     581..600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        799..833
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             842..853
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1596..1667
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1683..1710
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1830..2621,2622..2690))
                     /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     2453..2472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2691..2795)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaaaagtgcc acctgtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
       61 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      121 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      181 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      241 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      301 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      361 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      421 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      481 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      541 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      601 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      661 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      721 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg
      781 cggccgctcg gcgagacgtt gacggctagc tcagtcctag gtacagtgct agctactaga
      841 gaaagaggag aaatactaga tggtcagcaa aggcgaagaa gacaacatgg catcactgcc
      901 agcgacacac gaacttcata ttttcggctc catcaatggt gtggacttcg atatggttgg
      961 acagggtact gggaacccta acgatggata tgaggagttg aacttgaaga gtaccaaggg
     1021 cgatcttcag ttttcacctt ggattctcgt acctcatata ggttatggct ttcatcaata
     1081 tctgccctat cctgacggta tgtcaccttt ccaagcagct atggttgacg ggagcggtta
     1141 tcaggttcac cggacaatgc agtttgaaga tggagcctct ttgaccgtaa attatcggta
     1201 tacttatgag ggcagccata ttaaaggcga ggctcaagtt aaggggactg ggttccccgc
     1261 ggatggtccc gtaatgacaa acagtctgac ggctgcagac tggtgccgct ctaagaagac
     1321 atatccaaat gacaagacga ttattagcac tttcaagtgg tcatacacta caggaaatgg
     1381 taaacgctat aggagcactg cccggaccac gtatactttc gcgaagccga tggcggcaaa
     1441 ttatttgaaa aaccaaccga tgtatgtgtt cagaaagaca gaactcaagc attctaaaac
     1501 tgaactgaac ttcaaagagt ggcagaaagc ctttactgat gtaatgggca tggacgaact
     1561 ctataagtaa taaccaggca tactagtcca ggcatcaaat aaaacgaaag gctcagtcga
     1621 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     1681 ctggctcacc ttcgggtggg cctttctgcg tttatacgtc tctgaccgcg gccgcgatta
     1741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc
     1981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttcccc
//
