LOCUS       sequence-415844-        2986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid used for expressing his--tagged mNeongreen for protein 
            purification..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2986)
  AUTHORS   Meyerowitz JT, Larsson EM, Murray RM
  TITLE     Development of Cell-Free Transcription-Translation Systems in Three 
            Soil Pseudomonads.
  JOURNAL   ACS Synth Biol. 2024 Feb 16;13(2):530-537. doi: 
            10.1021/acssynbio.3c00468. Epub 2024 Feb 6.
  PUBMED    38319019
REFERENCE   2  (bases 1 to 2986)
  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/acssynbio.3c00468"; journalName: "ACS Synth Biol"; date: 
            "2024-02-16- 16"; volume: "13"; issue: "2"; pages: "530-537"
COMMENT     Sequence Label: pJTM-T7-mNeongreen-6xHis
FEATURES             Location/Qualifiers
     source          1..2986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(349..1140,1141..1209))
                     /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     972..991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1210..1314)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1382..1400
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1438..1460)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1621..1638)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2351..2373
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2380..2399)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2381..2399)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(join(2576..2986,1..178))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2656..2675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
       61 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      121 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      181 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      241 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      301 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      361 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      421 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      481 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      541 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      601 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      661 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      721 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      781 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      841 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      901 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      961 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1021 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1081 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1141 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1201 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1261 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1321 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     1381 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     1441 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggggt ggaagggctc
     1501 ggagttgtgg taatctatgt atcctggttg agaagagaaa agaaaaccgc cgatcctgtc
     1561 caccgcatta ctgcaaggta gtggacaaga ccggcggtct taagtttttt ggctgaatta
     1621 atggtgatgg tgatggtgct tatagagctc atccatcccc atcacatcgg taaacgcctt
     1681 ctgccattcc ttaaaattca gctccgtctt cgaatgcttg agctcggtct tccggaacac
     1741 atacatgggt tggttcttca gatagttcgc cgccatcggc ttggcaaagg tgtacgtcgt
     1801 gcgggccgtc gaacggtagc gcttgccgtt gccggtggta tacgaccact tgaacgtgct
     1861 gatgatggtc ttatcgttcg ggtacgtctt cttggaccga caccaatccg cggccgtcag
     1921 ggaattggtc atcaccggcc cgtccgccgg aaagccggtg cccttcacct gcgcttcccc
     1981 cttgatatgg ctgccttcgt acgtgtagcg atagttcacg gtcagggagg cgccatcctc
     2041 aaactgcatc gtgcgatgca cttggtaccc cgagccgtcg accatggccg cctgaaacgg
     2101 ggacatcccg tcggggtagg gcagatattg atggaagcca tacccgatgt gcggcaccag
     2161 gatccagggg ctaaactgca ggtcgccctt cgtgctcttg aggttgagct cttcgtagcc
     2221 atcattgggg ttgcccgtcc cttgccccac catatcaaag tccaccccgt tgatcgaccc
     2281 gaagatatgg agttcatgcg tcgcggggag ggacgccata ttatcttcct cgcccttgga
     2341 gaccattata tctccttctt aaagttaaac aaaattattc cctatagtga gtcgtattag
     2401 agacgagacg agacagcctg agaatggatg cgagtaatgt gcggcgagcg gtatcagctc
     2461 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2521 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2581 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2641 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2701 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2761 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2821 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2881 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2941 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaa
//
