LOCUS       sequence-234120-        6814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAGIC donor plasmid (Himar transposon with tetQ/sfGFP payload).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6814)
  AUTHORS   Ronda C, Chen SP, Cabral V, Yaung SJ, Wang HH
  TITLE     Metagenomic engineering of the mammalian gut microbiome in situ.
  JOURNAL   Nat Methods. 2019 Feb;16(2):167-170. doi: 10.1038/s41592-018-0301-y.
            Epub 2019 Jan 14.
  PUBMED    30643213
REFERENCE   2  (bases 1 to 6814)
  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: 
            "10.1038/s41592-018-0301-y"; journalName: "Nat Methods"; date: 
            "2019-02"; volume: "16"; issue: "2"; pages: "167-170"
COMMENT     Sequence Label: pGT-Ah9
FEATURES             Location/Qualifiers
     source          1..6814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..647
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     548..567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     764..781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            917..1026
                     /note="incP origin of transfer"
     CDS             complement(1347..2006)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1921..1940
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2007..2111)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2179..2197
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             4568..5281
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGFVTILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5341..5427
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5519..5546
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
       61 gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
      121 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
      181 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
      241 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
      301 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
      361 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
      421 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
      481 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
      541 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
      601 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
      661 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
      721 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
      781 ggaagcggaa tatatccaga tcctctacgc cggacgcatc gtggccggat caattccgtg
      841 ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt gccctcatct
      901 gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
      961 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
     1021 ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat
     1081 cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag
     1141 cagggttatg cagcggaaaa cggaattgat ccccctcgaa ttgacgcgta accagcaata
     1201 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga atttgctttc
     1261 gaatttctgc cattcatccg cttattatca cttattcagg cgtagcaacc aggcgtttaa
     1321 gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt
     1381 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     1441 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     1501 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     1561 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     1621 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     1681 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     1741 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgtaa ttccggatga
     1801 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     1861 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     1921 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     1981 gtatatccag tgattttttt ctccatactc ttcctttttc aatattattg aagcatttat
     2041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2101 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     2161 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcacgc gtcctcggta
     2221 cctaacaggt tggatgataa gtccccggtc ttcgtatgcc gtcttctgct tggcgcgccc
     2281 tcgagcaatt gccgaccgaa tttttatgtc gtaaagaggg gctttgcagg gggtggactc
     2341 agaaagatga gaatagatga ctattgtagt tgaaacacat agaaagttgc tgatatacag
     2401 accgatacgc atatcgggat gaaccatgag tacgttcttt tctcaaaaaa cataaatatt
     2461 cgaaaagaga tgcaataaat taaggagagg ttatactcta gagtagtaga ttattttagg
     2521 aatttagatg ttttgtatga aatagatgct tcgtatggaa ttaatgaaat ttttagtcag
     2581 gtaaaaaagg taataggaga atattatgaa tattataaat ttaggaattc ttgctcacat
     2641 tgatgcagga aaaacttccg taaccgagaa tctgctgttt gccagtggag caacggaaaa
     2701 gtgcggctgt gtggataatg gtgacaccat aacggactct atggatatag agaaacgtag
     2761 aggaattact gttcgggctt ctacgacatc tattatctgg aatggtgtga aatgcaatat
     2821 cattgacact ccgggacaca tggattttat tgcggaagtg gagcggacat tcaaaatgct
     2881 tgatggagca gtcctcatct tatccgcaaa ggaaggcata caagcgcaga caaagttgct
     2941 gttcaatact ttacagaagc tgcaaatccc gacaattata tttatcaata agattgaccg
     3001 agccggtgtg aatttggagc gtttgtatct ggatataaaa gcaaatctgt ctcaagatgt
     3061 cctgtttatg caaaatgttg tcgatggatc ggtttatccg gtttgctccc aaacatatat
     3121 aaaggaagaa tacaaagaat ttgtatgcaa ccatgacgac aatatattag aacgatattt
     3181 ggcggatagc gaaatttcac cggctgatta ttggaatacg ataatcgctc ttgtggcaaa
     3241 agccaaagtc tatccggtgc tacatggatc agcaatgttc aatatcggta tcaatgagtt
     3301 gttggacgcc atcacttctt ttatacttcc tccggcatcg gtctcaaaca gactttcatc
     3361 ttatctttat aagatagagc atgaccccaa aggacataaa agaagttttc taaaaataat
     3421 tgacggaagt ctgagacttc gagacgttgt aagaatcaac gattcggaaa aattcatcaa
     3481 gattaaaaat ctaaaaacta tcaatcaggg cagagagata aatgttgatg aagtgggcgc
     3541 caatgatatc gcgattgtag aggatatgga tgattttcga atcggaaatt atttaggtgc
     3601 tgaaccttgt ttgattcaag gattatcgca tcagcatccc gctctcaaat cctccgtccg
     3661 gccagacagg cccgaagaga gaagcaaggt gatatccgct ctgaatacat tgtggattga
     3721 agacccgtct ttgtcctttt ccataaactc atatagtgat gaattggaaa tctcgttata
     3781 tggtttaacc caaaaggaaa tcatacagac attgctggaa gaacgatttt ccgtaaaggt
     3841 ccattttgat gagatcaaga ctatatacaa agaacgacct gtaaaaaagg tcaataagat
     3901 tattcagatc gaagtgccgc ccaaccctta ttgggccaca atagggctga ctcttgaacc
     3961 cttaccgtta gggacagggt tgcaaatcga aagtgacatc tcctatggtt atctgaacca
     4021 ttcttttcaa aatgccgttt ttgaagggat tcgtatgtct tgccaatccg ggttacatgg
     4081 atgggaagtg actgatctga aagtaacttt tactcaagcc gagtattata gcccggtaag
     4141 tacacctgct gatttcagac agctgacccc ttatgtcttc aggctggcct tgcaacagtc
     4201 aggtgtggac attctcgaac cgatgctcta ttttgagttg cagatacccc aagcggcaag
     4261 ttccaaagct attacagatt tgcaaaaaat gatgtctgag attgaagaca tcagttgcaa
     4321 taatgagtgg tgtcatatta aagggaaagt tccattaaat acaagtaaag actatgcatc
     4381 agaagtaagt tcatacacta agggcttagg catttttatg gttaagccat gcgggtatca
     4441 aataacaaaa ggcggttatt ctgataatat ccgcatgaac gaaaaagata aacttttatt
     4501 catgttccaa aaatcaatgt catcaaaata aattatattt aaactcaatt taaagtaggg
     4561 agaatagatg cgtaaaggcg aagagctgtt cactggtttc gtcactattc tggtggaact
     4621 ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac
     4681 taatggtaaa ctgacgctga agttcatctg tactactggt aaactgccgg taccttggcc
     4741 gactctggta acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccacat
     4801 gaagcagcat gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat
     4861 ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac
     4921 cctggtaaac cgcattgagc tgaaaggcat tgactttaaa gaagacggca atatcctggg
     4981 ccataagctg gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa
     5041 aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct
     5101 ggctgatcac taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa
     5161 tcactatctg agcacgcaaa gcgttctgtc taaagatccg aacgagaaac gcgatcacat
     5221 ggttctgctg gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa
     5281 ataaaaacaa taggccacat gcaactgtct agaatgcgag agtagggaac tgccaggcat
     5341 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     5401 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     5461 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
     5521 aaggccatcc tgacggatgg cctttttgcg tttctacctg cagggcgcgc caagcagaag
     5581 acggcatacg aagaccgggg acttatcatc caacctgtta ggatccctct agagtagtag
     5641 attattttag gaatttagat gttttgtatg aaatagatgc ttcgtatgga attaatgaaa
     5701 tttttagtca ggtaaaaaag gtaataggag aatattcata tggaaaaaaa ggaatttcgt
     5761 gttttgataa aatactgttt tctgaaggga aaaaatacag tggaagcaaa aacttggctt
     5821 gataatgagt ttccggactc tgccccaggg aaatcaacaa taattgattg gtatgcaaaa
     5881 ttcaagcgtg gtgaaatgag cacggaggac ggtgaacgca gtggacgccc gaaagaggtg
     5941 gttaccgacg aaaacatcaa aaaaatccac aaaatgattt tgaatgaccg taaaatgaag
     6001 ttgatcgaga tagcagaggc cttaaagata tcaaaggaac gtgttggtca tatcattcat
     6061 caatatttgg atatgcggaa gctctgtgcg aaatgggtgc cgcgcgagct cacatttgac
     6121 caaaaacaac gacgtgttga tgattctaag cggtgtttgc agctgttaac tcgtaataca
     6181 cccgagtttt tccgtcgata tgtgacaatg gatgaaacat ggctccatca ctacactcct
     6241 gagtccaatc gacagtcggc tgagtggaca gcgaccggtg aaccgtctcc gaagcgtgga
     6301 aagactcaaa agtccgctgg caaagtaatg gcctctgttt tttgggatgc gcatggaata
     6361 atttttatcg attatcttga gaagggaaaa accatcaaca gtgactatta tatggcgtta
     6421 ttggagcgtt tgaaggtcga aatcgcggca aaacggcccc acatgaagaa gaaaaaagtg
     6481 ttgttccacc aagacaacgc accgtgccac aagtcattga gaacgatggc aaaaattcat
     6541 gaattgggct tcgaattgct tccccacccg ccgtattctc cagatctggc ccccagcgac
     6601 tttttcttgt tctcagacct caaaaggatg ctcgcaggga aaaaatttgg ctgcaatgaa
     6661 gaggtgatcg ccgaaactga ggcctatttt gaggcaaaac cgaaggagta ctaccaaaat
     6721 ggtatcaaaa aattggaagg tcgttataat cgttgtatcg ctcttgaagg gaactatgtt
     6781 gaataagcgg ccgccaccgc ggtggagggg aatt
//
