LOCUS       sequence-309669-        8359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aTc-inducible sfGFP expression on M. magneticum/E. coli shuttle 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8359)
  AUTHORS   Furubayashi M, Wallace AK, Gonzalez LM, Jahnke JP, Hanrahan BM, 
            Payne AL, Stratis-Cullum DN, Gray MT, Liu H, Rhoads MK, Voigt CA
  TITLE     Genetic Tuning of Iron Oxide Nanoparticle Size, Shape, and Surface 
            Properties in Magnetospirillum magneticum
  JOURNAL   Adv. Funct. Mater. 2021, 31, 2004813
REFERENCE   2  (bases 1 to 8359)
  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: "Adv. Funct.
            Mater. 2021, 31, 2004813"
COMMENT     Sequence Label: pMGA-ptet-sfGFP
FEATURES             Location/Qualifiers
     source          1..8359
                     /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"
     CDS             complement(141..764)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     672..691
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    952..970
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    977..995
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        1008..1057
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             1122..1835
                     /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      1887..1981
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      3906..3915
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      complement(5820..6365)
                     /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(5900..5919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6568..7362)
                     /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(6679..6698)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7289..7308
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     oriT            7758..7867
                     /note="incP origin of transfer"
     CDS             7900..8271
                     /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 tcaggaccca ctttcacatt taagttgttt ttctaatccg
      181 catatgatca attcaaggcc gaataagaag gctggctctg caccttggtg atcaaataat
      241 tcgatagctt gtcgtaataa tggcggcata ctatcagtag taggtgtttc cctttcttct
      301 ttagcgactt gatgctcttg atcttccaat acgcaaccta aagtaaaatg ccccacagcg
      361 ctgagtgcat ataatgcatt ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga
      421 ttttcgagag tttcatactg tttttctgta ggccgtgtac ctaaatgtac ttttgctcca
      481 tcgcgatgac ttagtaaagc acatctaaaa cttttagcgt tattacgtaa aaaatcttgc
      541 cagctttccc cttctaaagg gcaaaagtga gtatggtgcc tatctaacat ctcaatggct
      601 aaggcgtcga gcaaagcccg cttatttttt acatgccaat acaatgtagg ctgctctaca
      661 cctagcttct gggcgagttt acgggttgtt aaaccttcga ttccgacctc attaagcagc
      721 tctaatgcgc tgttaatcac tttactttta tctaatctgg acatattcac caccctgaat
      781 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggcg
      841 cgccgccatt ggaccaaaac gaaaaaaggc ccccctttcg ggaggcctct tttctggaat
      901 ttggtaccga gcttaacgat cgttggctgt tttcagcagg acgcactgac ctccctatca
      961 gtgatagaga ttgacatccc tatcagtgat agagatactg agcacagctg tcaccggatg
     1021 tgctttccgg tctgatgagt ccgtgaggac gaaacagcct ctacaaataa ttttgtttaa
     1081 tctagaaata attttgttta tctctcgagg aggtatacta gatgcgtaaa ggcgaagagc
     1141 tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac ggtcataagt
     1201 tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg ctgaagttca
     1261 tctgtactac tggtaaactg ccggtacctt ggccgactct ggtaacgacg ctgacttatg
     1321 gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc ttcaagtccg
     1381 ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac ggcacgtaca
     1441 aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt gagctgaaag
     1501 gcattgactt taaagaagac ggcaatatcc tgggccataa gctggaatac aattttaaca
     1561 gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg aattttaaaa
     1621 ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag caaaacactc
     1681 caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg caaagcgttc
     1741 tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc gtaaccgcag
     1801 cgggcatcac gcatggtatg gatgaactgt acaaatgaaa gcttgcggcc gcgtcgtgac
     1861 tgggaaaacc ctggcgacta gtcttggact cctgttgata gatccagtaa tgacctcaga
     1921 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
     1981 tccaggggtc cccaataatt acgatttaaa ttggcgggat ccaatcgctc gagcgcgagg
     2041 tcacaattgg agatgatccg ccagcgccga agccatcata actgcattga ataattcaaa
     2101 gtgcattgaa taaggcaggg tgcataaggc gcagcctttc gattaactcc atttcctgcg
     2161 gcgacaacgt cgccgcttgc ccccccgacg ggggtcaagc gaggcacagg gacccgccgc
     2221 ctcttggcgg tgggtggccg cagcttgacg cccgaacact ccgccgccca accctgcctg
     2281 ctctcctctg ttcaatgaaa ttgaacggag gagatgcctc ccggcgaggg cctttctttc
     2341 ttgcataacg ttcatgctac gttccaacgg ttataccgta tgagcgtcga actgtataat
     2401 cgttgaacag tcgaaccatc cccccgcgcc agcgcgggga gcagggatgg gttcgggggc
     2461 cggaagttag gcaaatcgcg gagccgtggc gccctcggta catccgcgaa atcggttgac
     2521 actttgcccc cagaacggct gcaacctacc gcttggcgac agccgcgtga cggtactttt
     2581 ccgctgtgga ttcggacacg aatatctgtt cagcagccag ggcgcgggcg atctggcgac
     2641 ggccgaggcc ctgatgggcc aattctgtga tccggtctat gaccgcttgg ctcatcggca
     2701 cccgaggacc gccgatacca gattggcgca catcgccaat ggtgggtgga accgagggag
     2761 ccggagccgg tgcagatttc tgcacagttg gttcacgccg tcgttctagc gcatcgagac
     2821 gctgcacgat cacggcgagc tgatcgttga cgtattgtcg ctcctcggac tgcgcaattg
     2881 tgatcagaaa ttcgaggagc tcggccacgg tgacgccgcg tcgaagggcc atagcctgta
     2941 tgtggtcgcg gaccgcatca ggacagcgaa tcgtccatct cgtgtctgac acattcccgg
     3001 tttttgtctg actcatcgtc gcccccaatg tctgacagat tcgagaaaga gtcataccgg
     3061 ctaacgcgaa cgctgacaac gaatgtctga caaaatgatg attacgtcag acattgatgt
     3121 ctgcctaaat cgcaatagtg tctgacatgg tttttggcgt tccgtatata aaatatgcgc
     3181 caaaatctag tgcgtcagcc aaacggcaaa cgcaaacagt tgacaccatt tccggtttta
     3241 tgagactgtg aaaagaccaa aggccgcctt gcggcggcct aggtgattgt ggtcccggcg
     3301 accaaggctg cgaaaacttg gcggtccagc agcagaggaa gcccgtttga aaggtccggc
     3361 ttgcgccgga accacctaat gacaatagca attcgccgca cagttttcaa cacccagccc
     3421 gccatgattc gcggcggcgt tgaacgttga acaacagacg tctcaaacag acacccgcaa
     3481 acgcgaacgg tatcggttgc aggatgcagc cgctgaactg ctgccggatt atcgggtgcg
     3541 gcattgccac cgctcaattg gatgtggctg tggcgagttg gtcagcgttg ttaccaatgg
     3601 gcagtcggcc aatttcaaag gcgtgcagac ctgcggctcg gtctggcact gcccaatctg
     3661 ttcggcaaaa atcagcaatg ttcggcgcgg cgagctgaac gagctgctgc gctgggggcg
     3721 ggagagggag ctgatcccgc tgatggtgac gctgacaggc catcacaatg cacggatgag
     3781 cctcgctgaa tgtctgacgg tgatgaagaa ggccaagcag agccttcacc agtctggccc
     3841 atggaagcgc catgttggcc cggtggtcga agggcacgtc acggcaacgg aggtcaatcg
     3901 gtcacgccgc catggctggc acgtccattt tcacgtcctg atgctgattg aggccgccag
     3961 cgaggccgaa gcgattgcgc tcgtgacccg acacctcgaa cctgaatggc tccggcagct
     4021 tgagcgccaa ggcagctggg gcggcgaaca tgcgttagac gtccagggcg cagccgatgc
     4081 ggatcggtat gtggccaagt ggggcgcggc tgaggaattg gctctgtcgg gcgaaaagcg
     4141 ggcgcgcggc gacgccgaaa ttaagggtca aacgccgtgg gaattgttgg ccgctgcggc
     4201 tgatggtgac gtccaggctg gccgcgattt tgtggagtac gcgaccctgt ttcatggcaa
     4261 acgccagctc gtttggtcgc gaggactgaa agagcgtgtc ggcctcaagg tggtggaaga
     4321 cgacgaggcc gccggcgagc tggaggatgt cgcagatagc gccatcaacg aagtggccgt
     4381 gttgccgagg ccgacttggc gtcacatcgt ccggcgcagg ctccgcgcgg atctgctggc
     4441 cgccgctcgt gatgacggat tcgatggcgt cgcgcggctg ctcaatgagc atggcatcgg
     4501 cggagctatg cccggcggct ggcgaccaat gcttagcgaa tataccgagc atattgcgat
     4561 ggctgcgtga ctcttttggc ccaccggcct gagggaaatt tgcctcgcag agctcacgat
     4621 aatttgacaa gggccgtagg cccgcaggca aattgtctag tgagtgtaca attttcaaat
     4681 tcgaaggtta ttgtagagct gacgctctca aggtgatgac gcctatggct gacgattctg
     4741 gcgtctggcg gattgtttgt cggattagca aaatccacac gatgggcgtg ctgaaggccc
     4801 ggtcgaatca caatctccga acgccaggtc atgaccagct cgcacatgtt gaccaggcca
     4861 aatccgctca gaatgaaatt ctgattggca ccggcgacgc catgctcgat gtgcaatcag
     4921 cattgaaatc gcggctggtc ggcaacattc gaaaaaatgg cgttatcgca gtggaaatgg
     4981 tgatgacagc ccgggcggac tggttcgctg ccgatcccgc gcgagctgcc gcatttaagc
     5041 agcgggtgct ggacgaggcg aatgccagat atggggcgaa tctcgtaacc gcagtgtggc
     5101 acatggatga agaggctccc catctgcatc tgtatatcgt gccgatggac caacggggca
     5161 aactgaactg ccgcgcgcag ttctcgacga tggcgtcgct gaaacaactc caggattggg
     5221 ctggcgtggt cggcaatcca ctcggcctcg aacgcggacg gccaaaagtg gagtttattg
     5281 tcgatggcga tgagccgccg gagaaccaaa gcccggcgga atatcgagct caaaccaaac
     5341 gcgacgctga agcggcagcc gccgcgcgga tcgctgccga gcgtgatgct gccgaggctg
     5401 cgcgactgct gattgaggct caggagcagg ccgccgcgat ccgcgcggcc caggctgaaa
     5461 ttatggcaga gcgtcgccag ctcaatgagg ataaggcgag ttttgctgag acgctgcgta
     5521 gccacacgag ggcgatgggc ctggcgtttc gttcatggct ggccaatttc gtcggccgcg
     5581 gccagatgca ggctgatgga gggccattgc tgacctttaa agccgcatgc cccgccagta
     5641 agcgagagga gctacagtcc gagccagcta atgcgtacgc gcgcctggtg gtcgctcata
     5701 tgcctgatcg tgacgctctt gccgtgctcg aaaccaaaag cgcggaggcc aaggaatagc
     5761 ttggcgtaat catgggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
     5821 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     5881 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
     5941 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     6001 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     6061 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6121 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     6181 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     6241 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
     6301 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     6361 ctcaagaaga tcatcttatt aatcagataa aatatttcaa gatttagaac tccagcatga
     6421 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     6481 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     6541 cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     6601 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     6661 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     6721 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     6781 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt
     6841 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     6901 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     6961 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     7021 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     7081 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     7141 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc
     7201 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     7261 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     7321 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
     7381 tgtctcttga tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat
     7441 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg
     7501 ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc
     7561 tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagg ggaagcgtgg
     7621 cgcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca
     7681 tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt
     7741 atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc
     7801 ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc
     7861 tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga aaccaagcca
     7921 accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg aagagcgatt
     7981 gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc cgtcggccag
     8041 ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc ccgcatcaat
     8101 ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga cccgcgcacg
     8161 gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga gaagcaggac
     8221 gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg acttttttag
     8281 ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc gctccataca
     8341 gttagggcgc gcccagctg
//
