LOCUS       sequence-309647-        7469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M. magneticum/E. coli shuttle vector (pMGT and p15A ori).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7469)
  AUTHORS   Schuerle S, Furubayashi M, Soleimany AP, Gwisai T, Huang W, Voigt C,
            Bhatia SN
  TITLE     Genetic Encoding of Targeted Magnetic Resonance Imaging Contrast 
            Agents for Tumor Imaging.
  JOURNAL   ACS Synth Biol. 2020 Feb 21;9(2):392-401. doi: 
            10.1021/acssynbio.9b00416. Epub 2020 Jan 22.
  PUBMED    31922737
REFERENCE   2  (bases 1 to 7469)
  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.9b00416"; journalName: "ACS Synth Biol"; date: 
            "2020-02-21- 21"; volume: "9"; issue: "2"; pages: "392-401"
COMMENT     Sequence Label: pMGA
FEATURES             Location/Qualifiers
     source          1..7469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(14..100)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        148..182
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             224..937
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      997..1091
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      3016..3025
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      complement(4930..5475)
                     /direction=LEFT
                     /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     complement(5010..5029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5678..6472)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5789..5808)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6399..6418
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     oriT            6868..6977
                     /note="incP origin of transfer"
     CDS             7010..7381
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 tctagggcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
       61 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctttaa ttaaagcgga
      121 taacaatttc agaattctca tgagaccttg acagctagct cagtcctagg tataatgcta
      181 gctctagaaa taattttgtt tatctctcga ggaggtatac tagatgcgta aaggcgaaga
      241 gctgttcact ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca acggtcataa
      301 gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt
      361 catctgtact actggtaaac tgccggtacc ttggccgact ctggtaacga cgctgactta
      421 tggtgttcag tgctttgctc gttatccgga ccatatgaag cagcatgact tcttcaagtc
      481 cgccatgccg gaaggctatg tgcaggaacg cacgatttcc tttaaggatg acggcacgta
      541 caaaacgcgt gcggaagtga aatttgaagg cgataccctg gtaaaccgca ttgagctgaa
      601 aggcattgac tttaaagaag acggcaatat cctgggccat aagctggaat acaattttaa
      661 cagccacaat gtttacatca ccgccgataa acaaaaaaat ggcattaaag cgaattttaa
      721 aattcgccac aacgtggagg atggcagcgt gcagctggct gatcactacc agcaaaacac
      781 tccaatcggt gatggtcctg ttctgctgcc agacaatcac tatctgagca cgcaaagcgt
      841 tctgtctaaa gatccgaacg agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc
      901 agcgggcatc acgcatggta tggatgaact gtacaaatga tggtctcaaa gcttgcggcc
      961 gcgtcgtgac tgggaaaacc ctggcgacta gtcttggact cctgttgata gatccagtaa
     1021 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     1081 ttggtgagaa tccaggggtc cccaataatt acgatttaaa ttggcgggat ccaatcgctc
     1141 gagcgcgagg tcacaattgg agatgatccg ccagcgccga agccatcata actgcattga
     1201 ataattcaaa gtgcattgaa taaggcaggg tgcataaggc gcagcctttc gattaactcc
     1261 atttcctgcg gcgacaacgt cgccgcttgc ccccccgacg ggggtcaagc gaggcacagg
     1321 gacccgccgc ctcttggcgg tgggtggccg cagcttgacg cccgaacact ccgccgccca
     1381 accctgcctg ctctcctctg ttcaatgaaa ttgaacggag gagatgcctc ccggcgaggg
     1441 cctttctttc ttgcataacg ttcatgctac gttccaacgg ttataccgta tgagcgtcga
     1501 actgtataat cgttgaacag tcgaaccatc cccccgcgcc agcgcgggga gcagggatgg
     1561 gttcgggggc cggaagttag gcaaatcgcg gagccgtggc gccctcggta catccgcgaa
     1621 atcggttgac actttgcccc cagaacggct gcaacctacc gcttggcgac agccgcgtga
     1681 cggtactttt ccgctgtgga ttcggacacg aatatctgtt cagcagccag ggcgcgggcg
     1741 atctggcgac ggccgaggcc ctgatgggcc aattctgtga tccggtctat gaccgcttgg
     1801 ctcatcggca cccgaggacc gccgatacca gattggcgca catcgccaat ggtgggtgga
     1861 accgagggag ccggagccgg tgcagatttc tgcacagttg gttcacgccg tcgttctagc
     1921 gcatcgagac gctgcacgat cacggcgagc tgatcgttga cgtattgtcg ctcctcggac
     1981 tgcgcaattg tgatcagaaa ttcgaggagc tcggccacgg tgacgccgcg tcgaagggcc
     2041 atagcctgta tgtggtcgcg gaccgcatca ggacagcgaa tcgtccatct cgtgtctgac
     2101 acattcccgg tttttgtctg actcatcgtc gcccccaatg tctgacagat tcgagaaaga
     2161 gtcataccgg ctaacgcgaa cgctgacaac gaatgtctga caaaatgatg attacgtcag
     2221 acattgatgt ctgcctaaat cgcaatagtg tctgacatgg tttttggcgt tccgtatata
     2281 aaatatgcgc caaaatctag tgcgtcagcc aaacggcaaa cgcaaacagt tgacaccatt
     2341 tccggtttta tgagactgtg aaaagaccaa aggccgcctt gcggcggcct aggtgattgt
     2401 ggtcccggcg accaaggctg cgaaaacttg gcggtccagc agcagaggaa gcccgtttga
     2461 aaggtccggc ttgcgccgga accacctaat gacaatagca attcgccgca cagttttcaa
     2521 cacccagccc gccatgattc gcggcggcgt tgaacgttga acaacagacg tctcaaacag
     2581 acacccgcaa acgcgaacgg tatcggttgc aggatgcagc cgctgaactg ctgccggatt
     2641 atcgggtgcg gcattgccac cgctcaattg gatgtggctg tggcgagttg gtcagcgttg
     2701 ttaccaatgg gcagtcggcc aatttcaaag gcgtgcagac ctgcggctcg gtctggcact
     2761 gcccaatctg ttcggcaaaa atcagcaatg ttcggcgcgg cgagctgaac gagctgctgc
     2821 gctgggggcg ggagagggag ctgatcccgc tgatggtgac gctgacaggc catcacaatg
     2881 cacggatgag cctcgctgaa tgtctgacgg tgatgaagaa ggccaagcag agccttcacc
     2941 agtctggccc atggaagcgc catgttggcc cggtggtcga agggcacgtc acggcaacgg
     3001 aggtcaatcg gtcacgccgc catggctggc acgtccattt tcacgtcctg atgctgattg
     3061 aggccgccag cgaggccgaa gcgattgcgc tcgtgacccg acacctcgaa cctgaatggc
     3121 tccggcagct tgagcgccaa ggcagctggg gcggcgaaca tgcgttagac gtccagggcg
     3181 cagccgatgc ggatcggtat gtggccaagt ggggcgcggc tgaggaattg gctctgtcgg
     3241 gcgaaaagcg ggcgcgcggc gacgccgaaa ttaagggtca aacgccgtgg gaattgttgg
     3301 ccgctgcggc tgatggtgac gtccaggctg gccgcgattt tgtggagtac gcgaccctgt
     3361 ttcatggcaa acgccagctc gtttggtcgc gaggactgaa agagcgtgtc ggcctcaagg
     3421 tggtggaaga cgacgaggcc gccggcgagc tggaggatgt cgcagatagc gccatcaacg
     3481 aagtggccgt gttgccgagg ccgacttggc gtcacatcgt ccggcgcagg ctccgcgcgg
     3541 atctgctggc cgccgctcgt gatgacggat tcgatggcgt cgcgcggctg ctcaatgagc
     3601 atggcatcgg cggagctatg cccggcggct ggcgaccaat gcttagcgaa tataccgagc
     3661 atattgcgat ggctgcgtga ctcttttggc ccaccggcct gagggaaatt tgcctcgcag
     3721 agctcacgat aatttgacaa gggccgtagg cccgcaggca aattgtctag tgagtgtaca
     3781 attttcaaat tcgaaggtta ttgtagagct gacgctctca aggtgatgac gcctatggct
     3841 gacgattctg gcgtctggcg gattgtttgt cggattagca aaatccacac gatgggcgtg
     3901 ctgaaggccc ggtcgaatca caatctccga acgccaggtc atgaccagct cgcacatgtt
     3961 gaccaggcca aatccgctca gaatgaaatt ctgattggca ccggcgacgc catgctcgat
     4021 gtgcaatcag cattgaaatc gcggctggtc ggcaacattc gaaaaaatgg cgttatcgca
     4081 gtggaaatgg tgatgacagc ccgggcggac tggttcgctg ccgatcccgc gcgagctgcc
     4141 gcatttaagc agcgggtgct ggacgaggcg aatgccagat atggggcgaa tctcgtaacc
     4201 gcagtgtggc acatggatga agaggctccc catctgcatc tgtatatcgt gccgatggac
     4261 caacggggca aactgaactg ccgcgcgcag ttctcgacga tggcgtcgct gaaacaactc
     4321 caggattggg ctggcgtggt cggcaatcca ctcggcctcg aacgcggacg gccaaaagtg
     4381 gagtttattg tcgatggcga tgagccgccg gagaaccaaa gcccggcgga atatcgagct
     4441 caaaccaaac gcgacgctga agcggcagcc gccgcgcgga tcgctgccga gcgtgatgct
     4501 gccgaggctg cgcgactgct gattgaggct caggagcagg ccgccgcgat ccgcgcggcc
     4561 caggctgaaa ttatggcaga gcgtcgccag ctcaatgagg ataaggcgag ttttgctgag
     4621 acgctgcgta gccacacgag ggcgatgggc ctggcgtttc gttcatggct ggccaatttc
     4681 gtcggccgcg gccagatgca ggctgatgga gggccattgc tgacctttaa agccgcatgc
     4741 cccgccagta agcgagagga gctacagtcc gagccagcta atgcgtacgc gcgcctggtg
     4801 gtcgctcata tgcctgatcg tgacgctctt gccgtgctcg aaaccaaaag cgcggaggcc
     4861 aaggaatagc ttggcgtaat catgggaaga tacttaacag ggaagtgaga gggccgcggc
     4921 aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa
     4981 tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc
     5041 cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc
     5101 gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg
     5161 actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
     5221 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
     5281 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
     5341 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
     5401 ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
     5461 ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcaa gatttagaac
     5521 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     5581 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     5641 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     5701 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     5761 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     5821 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     5881 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     5941 tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     6001 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     6061 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     6121 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     6181 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     6241 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat
     6301 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     6361 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     6421 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     6481 atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc cttggcggca
     6541 agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg
     6601 gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc
     6661 cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagg
     6721 ggaagcgtgg cgcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc
     6781 ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt
     6841 tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag
     6901 cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc
     6961 tctgcgaggc tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga
     7021 aaccaagcca accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg
     7081 aagagcgatt gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc
     7141 cgtcggccag ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc
     7201 ccgcatcaat ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga
     7261 cccgcgcacg gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga
     7321 gaagcaggac gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg
     7381 acttttttag ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc
     7441 gctccataca gttagggcgc gcccagctg
//
