LOCUS       sequence-309668-        8800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-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 8800)
  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 8800)
  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-ptac-sfGFP
FEATURES             Location/Qualifiers
     source          1..8800
                     /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(146..1228)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYMTVSRVVNQASHVSAKTREKVEAAMVEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIYSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1190..1209
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1393..1398,1399..1414,1415..1421)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1427..1446
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     misc_RNA        1449..1498
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             1563..2276
                     /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      2328..2422
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      4347..4356
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      complement(6261..6806)
                     /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(6341..6360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7009..7803)
                     /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(7120..7139)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7730..7749
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     oriT            8199..8308
                     /note="incP origin of transfer"
     CDS             8341..8712
                     /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 ccttatcact gcccgctttc cagtcgggaa acctgtcgtg
      181 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
      241 gggtggtttt tcttttcacc agtgagactg gcaacagctg attgcccttc accgcctggc
      301 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
      361 tgatggtggt taacggcggg atataacatg agctatcttc ggtatcgtcg tatcccacta
      421 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
      481 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
      541 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
      601 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
      661 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
      721 ccagtcgcgt accgtcctca tgggagtaaa taatactgtt gatgggtgtc tggtcagaga
      781 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
      841 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
      901 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
      961 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     1021 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     1081 tgggaatgta attcagctcc accatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     1141 cgtggctggc ctggttcacc acgcgggaaa cggtcatata agagacaccg gcatactctg
     1201 cgacatcgta taacgttact ggtttcatat tcaccaccct gaattgactc tcttccgggc
     1261 gctatcatgc cataccgcga aaggttttgc gccattcgat ggcgcgccgc cattggacca
     1321 aaacgaaaaa aggcccccct ttcgggaggc ctcttttctg gaatttggta ccgagaacga
     1381 tcgttggctg tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcgctc
     1441 acaattagct gtcaccggat gtgctttccg gtctgatgag tccgtgagga cgaaacagcc
     1501 tctacaaata attttgttta atctagaaat aattttgttt atctctcgag gaggtatact
     1561 agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc tattctggtg gaactggatg
     1621 gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg
     1681 gtaaactgac gctgaagttc atctgtacta ctggtaaact gccggtacct tggccgactc
     1741 tggtaacgac gctgacttat ggtgttcagt gctttgctcg ttatccggac catatgaagc
     1801 agcatgactt cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc acgatttcct
     1861 ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg
     1921 taaaccgcat tgagctgaaa ggcattgact ttaaagaaga cggcaatatc ctgggccata
     1981 agctggaata caattttaac agccacaatg tttacatcac cgccgataaa caaaaaaatg
     2041 gcattaaagc gaattttaaa attcgccaca acgtggagga tggcagcgtg cagctggctg
     2101 atcactacca gcaaaacact ccaatcggtg atggtcctgt tctgctgcca gacaatcact
     2161 atctgagcac gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat catatggttc
     2221 tgctggagtt cgtaaccgca gcgggcatca cgcatggtat ggatgaactg tacaaatgaa
     2281 agcttgcggc cgcgtcgtga ctgggaaaac cctggcgact agtcttggac tcctgttgat
     2341 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     2401 ggcgtttttt attggtgaga atccaggggt ccccaataat tacgatttaa attggcggga
     2461 tccaatcgct cgagcgcgag gtcacaattg gagatgatcc gccagcgccg aagccatcat
     2521 aactgcattg aataattcaa agtgcattga ataaggcagg gtgcataagg cgcagccttt
     2581 cgattaactc catttcctgc ggcgacaacg tcgccgcttg cccccccgac gggggtcaag
     2641 cgaggcacag ggacccgccg cctcttggcg gtgggtggcc gcagcttgac gcccgaacac
     2701 tccgccgccc aaccctgcct gctctcctct gttcaatgaa attgaacgga ggagatgcct
     2761 cccggcgagg gcctttcttt cttgcataac gttcatgcta cgttccaacg gttataccgt
     2821 atgagcgtcg aactgtataa tcgttgaaca gtcgaaccat ccccccgcgc cagcgcgggg
     2881 agcagggatg ggttcggggg ccggaagtta ggcaaatcgc ggagccgtgg cgccctcggt
     2941 acatccgcga aatcggttga cactttgccc ccagaacggc tgcaacctac cgcttggcga
     3001 cagccgcgtg acggtacttt tccgctgtgg attcggacac gaatatctgt tcagcagcca
     3061 gggcgcgggc gatctggcga cggccgaggc cctgatgggc caattctgtg atccggtcta
     3121 tgaccgcttg gctcatcggc acccgaggac cgccgatacc agattggcgc acatcgccaa
     3181 tggtgggtgg aaccgaggga gccggagccg gtgcagattt ctgcacagtt ggttcacgcc
     3241 gtcgttctag cgcatcgaga cgctgcacga tcacggcgag ctgatcgttg acgtattgtc
     3301 gctcctcgga ctgcgcaatt gtgatcagaa attcgaggag ctcggccacg gtgacgccgc
     3361 gtcgaagggc catagcctgt atgtggtcgc ggaccgcatc aggacagcga atcgtccatc
     3421 tcgtgtctga cacattcccg gtttttgtct gactcatcgt cgcccccaat gtctgacaga
     3481 ttcgagaaag agtcataccg gctaacgcga acgctgacaa cgaatgtctg acaaaatgat
     3541 gattacgtca gacattgatg tctgcctaaa tcgcaatagt gtctgacatg gtttttggcg
     3601 ttccgtatat aaaatatgcg ccaaaatcta gtgcgtcagc caaacggcaa acgcaaacag
     3661 ttgacaccat ttccggtttt atgagactgt gaaaagacca aaggccgcct tgcggcggcc
     3721 taggtgattg tggtcccggc gaccaaggct gcgaaaactt ggcggtccag cagcagagga
     3781 agcccgtttg aaaggtccgg cttgcgccgg aaccacctaa tgacaatagc aattcgccgc
     3841 acagttttca acacccagcc cgccatgatt cgcggcggcg ttgaacgttg aacaacagac
     3901 gtctcaaaca gacacccgca aacgcgaacg gtatcggttg caggatgcag ccgctgaact
     3961 gctgccggat tatcgggtgc ggcattgcca ccgctcaatt ggatgtggct gtggcgagtt
     4021 ggtcagcgtt gttaccaatg ggcagtcggc caatttcaaa ggcgtgcaga cctgcggctc
     4081 ggtctggcac tgcccaatct gttcggcaaa aatcagcaat gttcggcgcg gcgagctgaa
     4141 cgagctgctg cgctgggggc gggagaggga gctgatcccg ctgatggtga cgctgacagg
     4201 ccatcacaat gcacggatga gcctcgctga atgtctgacg gtgatgaaga aggccaagca
     4261 gagccttcac cagtctggcc catggaagcg ccatgttggc ccggtggtcg aagggcacgt
     4321 cacggcaacg gaggtcaatc ggtcacgccg ccatggctgg cacgtccatt ttcacgtcct
     4381 gatgctgatt gaggccgcca gcgaggccga agcgattgcg ctcgtgaccc gacacctcga
     4441 acctgaatgg ctccggcagc ttgagcgcca aggcagctgg ggcggcgaac atgcgttaga
     4501 cgtccagggc gcagccgatg cggatcggta tgtggccaag tggggcgcgg ctgaggaatt
     4561 ggctctgtcg ggcgaaaagc gggcgcgcgg cgacgccgaa attaagggtc aaacgccgtg
     4621 ggaattgttg gccgctgcgg ctgatggtga cgtccaggct ggccgcgatt ttgtggagta
     4681 cgcgaccctg tttcatggca aacgccagct cgtttggtcg cgaggactga aagagcgtgt
     4741 cggcctcaag gtggtggaag acgacgaggc cgccggcgag ctggaggatg tcgcagatag
     4801 cgccatcaac gaagtggccg tgttgccgag gccgacttgg cgtcacatcg tccggcgcag
     4861 gctccgcgcg gatctgctgg ccgccgctcg tgatgacgga ttcgatggcg tcgcgcggct
     4921 gctcaatgag catggcatcg gcggagctat gcccggcggc tggcgaccaa tgcttagcga
     4981 atataccgag catattgcga tggctgcgtg actcttttgg cccaccggcc tgagggaaat
     5041 ttgcctcgca gagctcacga taatttgaca agggccgtag gcccgcaggc aaattgtcta
     5101 gtgagtgtac aattttcaaa ttcgaaggtt attgtagagc tgacgctctc aaggtgatga
     5161 cgcctatggc tgacgattct ggcgtctggc ggattgtttg tcggattagc aaaatccaca
     5221 cgatgggcgt gctgaaggcc cggtcgaatc acaatctccg aacgccaggt catgaccagc
     5281 tcgcacatgt tgaccaggcc aaatccgctc agaatgaaat tctgattggc accggcgacg
     5341 ccatgctcga tgtgcaatca gcattgaaat cgcggctggt cggcaacatt cgaaaaaatg
     5401 gcgttatcgc agtggaaatg gtgatgacag cccgggcgga ctggttcgct gccgatcccg
     5461 cgcgagctgc cgcatttaag cagcgggtgc tggacgaggc gaatgccaga tatggggcga
     5521 atctcgtaac cgcagtgtgg cacatggatg aagaggctcc ccatctgcat ctgtatatcg
     5581 tgccgatgga ccaacggggc aaactgaact gccgcgcgca gttctcgacg atggcgtcgc
     5641 tgaaacaact ccaggattgg gctggcgtgg tcggcaatcc actcggcctc gaacgcggac
     5701 ggccaaaagt ggagtttatt gtcgatggcg atgagccgcc ggagaaccaa agcccggcgg
     5761 aatatcgagc tcaaaccaaa cgcgacgctg aagcggcagc cgccgcgcgg atcgctgccg
     5821 agcgtgatgc tgccgaggct gcgcgactgc tgattgaggc tcaggagcag gccgccgcga
     5881 tccgcgcggc ccaggctgaa attatggcag agcgtcgcca gctcaatgag gataaggcga
     5941 gttttgctga gacgctgcgt agccacacga gggcgatggg cctggcgttt cgttcatggc
     6001 tggccaattt cgtcggccgc ggccagatgc aggctgatgg agggccattg ctgaccttta
     6061 aagccgcatg ccccgccagt aagcgagagg agctacagtc cgagccagct aatgcgtacg
     6121 cgcgcctggt ggtcgctcat atgcctgatc gtgacgctct tgccgtgctc gaaaccaaaa
     6181 gcgcggaggc caaggaatag cttggcgtaa tcatgggaag atacttaaca gggaagtgag
     6241 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     6301 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6361 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     6421 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     6481 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     6541 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     6601 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     6661 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     6721 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     6781 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttca
     6841 agatttagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
     6901 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
     6961 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
     7021 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     7081 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     7141 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     7201 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     7261 gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     7321 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     7381 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     7441 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     7501 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     7561 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     7621 ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     7681 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     7741 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     7801 catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc gccatcagat
     7861 ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg
     7921 cgccccagct ggcaattccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg
     7981 ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca
     8041 gatagcccag gggaagcgtg gcgcttttcc gctgcataac cctgcttcgg ggtcattata
     8101 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
     8161 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     8221 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     8281 gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg
     8341 atggctgatg aaaccaagcc aaccaggaag ggcagcccac ctatcaaggt gtactgcctt
     8401 ccagacgaac gaagagcgat tgaggaaaag gcggcggcgg ccggcatgag cctgtcggcc
     8461 tacctgctgg ccgtcggcca gggctacaaa atcacgggcg tcgtggacta tgagcacgtc
     8521 cgcgagctgg cccgcatcaa tggcgacctg ggccgcctgg gcggcctgct gaaactctgg
     8581 ctcaccgacg acccgcgcac ggcgcggttc ggtgatgcca cgatcctcgc cctgctggcg
     8641 aagatcgaag agaagcagga cgagcttggc aaggtcatga tgggcgtggt ccgcccgagg
     8701 gcagagccat gactttttta gccgctaaaa cggccggggg gtgcgcgtga ttgccaagca
     8761 cgtccccatg cgctccatac agttagggcg cgcccagctg
//
