LOCUS       sequence-387898-        6869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Second-generation mammalian acoustic reporter gene derived from 
            Anabaena.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6869)
  AUTHORS   Hurt RC, Buss MT, Duan M, Wong K, You MY, Sawyer DP, Swift MB, Dutka
            P, Barturen-Larrea P, Mittelstein DR, Jin Z, Abedi MH, Farhadi A, 
            Deshpande R, Shapiro MG
  TITLE     Genomically mined acoustic reporter genes for real-time in vivo 
            monitoring of tumors and tumor-homing bacteria.
  JOURNAL   Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01581-y.
  PUBMED    36593411
REFERENCE   2  (bases 1 to 6869)
  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: "Nat 
            Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01581-y."
COMMENT     Sequence Label: mARGAna (cassette 1, transient)
FEATURES             Location/Qualifiers
     source          1..6869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    24..40
                     /label=lac operator
                     /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)."
     primer_bind     29..51
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     enhancer        70..373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        374..577
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     528..548
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     574..598
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        693..711
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     693..710
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    1006..1557
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1150..1167)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1377..1396
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(2342..2361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2343..2361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2756..3344
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2809..2829)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3227..3238)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3364..3840
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3563..3582)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      3976..4430
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4063..4082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4272..4293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(4717..4785,4786..5577)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAVLGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4935..4954)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    5679..5712
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      5790..6378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6279..6298
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6547..6564
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(6638..6742)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6810..6828
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(6855..6860,6861..6869,1..9,10..16)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
ORIGIN
        1 tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcat
       61 cgtgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      121 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      181 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      241 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      301 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      361 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      421 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      481 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      541 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
      601 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
      661 ccggactcta gcctaggctt ttgcaaaaag ctatttaggt gacactatag aaggtacgcc
      721 tgcaggtacc gagctcggat ccagtaccct tcaccatggc cgtggaaaag accaacagca
      781 gcagctccct ggccgaagtg atcgacagaa tcctggacaa gggcatcgtg atcgacgcct
      841 gggtgcgcgt gtccctcgtg ggaattgagc tgctggccat cgaggcccgg atcgtgattg
      901 ccagcgtgga aacctacctg aagtacgccg aggccgtggg cctgacacag agtgctgctg
      961 tgcctgcttg ataatactaa atcgcactgt cggcgtcccc ccctaacgtt actggccgaa
     1021 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     1081 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     1141 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     1201 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     1261 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     1321 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     1381 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     1441 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg
     1501 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg
     1561 ccacaaccat ggtgtccaag ggcgaggaac tgttcaccgg cgtggtgccc atcctggtgg
     1621 aactggatgg cgacgtgaac ggccacaagt tctctgtgcg gggagagggc gaaggcgacg
     1681 ccacaaatgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgcctt
     1741 ggcctaccct cgtgaccaca ctgtctcacg gcgtgcagtg cttcgccaga taccccgacc
     1801 acatgaagca gcacgatttc ttcaagagcg ccatgcccga gggctacgtg caggaacgga
     1861 ccatcttctt caaggacgac ggcacctaca agaccagagc cgaagtgaag ttcgagggcg
     1921 acaccctcgt gaaccggatc gagctgaagg gcgtggactt caaagaggac ggcaacatcc
     1981 tgggccacaa gctggagtac aacttcaaca gccacaacat ctacatcatg gccgtgaagc
     2041 agaaaaacgg catcaaagtg aacttcaaga tccggcacaa cgtggaagat ggcagcgtgc
     2101 agctggccga ccactaccag cagaacaccc ccatcggaga tggccccgtg ctgctgcctg
     2161 atagccacta cctgagcacc cagagcgtgc tgagcaagga ccccaacgag aagcgggacc
     2221 acatggtgct gctggaattt cggaccgccg ctggcatcac cctgggcatg gatgagctgt
     2281 acaagtgaag gatccctcga ggggcccaag cttacgcgtg catgcgacgt catagctctc
     2341 tccctatagt gagtcgtatt ataagctagc ttgggatctt tgtgaaggaa ccttacttct
     2401 gtggtgtgac ataattggac aaactaccta cagagattta aagctctaag gtaaatataa
     2461 aatttttaag tgtataatgt gttaaactag ctgcatatgc ttgctgcttg agagttttgc
     2521 ttactgagta tgatttatga aaatattata cacaggagct agtgattcta attgtttgtg
     2581 tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac agtctgttca
     2641 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2701 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttcgt ggctgaatca
     2761 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2821 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2881 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2941 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3001 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3061 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3121 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     3181 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3241 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3301 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgccagaga caattaactt
     3361 cgcgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     3421 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     3481 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     3541 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     3601 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     3661 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     3721 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     3781 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     3841 gaagttctca gatcctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3901 ttggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3961 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4021 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4081 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4141 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4201 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4261 cgttcttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4321 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4381 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     4441 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4501 tatccgctca tgccaggtct tggactggtg agaacggctt gctcggcagc ttcgatgtgt
     4561 gctggaggga gaataaaggt ctaagatgtg cgatagaggg aagtcgcatt gaattatgtg
     4621 ctgtgtaggg atcgctggta tcaaatatgt gtgcccaccc ctggcatgag acaataaccc
     4681 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4741 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4801 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4861 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4921 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4981 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5041 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5101 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5161 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5221 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5281 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5341 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5401 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agtactgggg
     5461 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5521 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5581 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5641 aggatctagg tgaagatcct ttttgataat ctcatgccat aacttcgtat aatgtatgct
     5701 atacgaagtt atggcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5761 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5821 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5881 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5941 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6001 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6061 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6121 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6181 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6241 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6301 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6361 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6421 tggccttttg ctcacatgtt ctttcctgcg ttattctttc ctgcgttatc ccctgattct
     6481 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6541 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6601 cccgcgcgtt ggccgattca ttaatgcaga gcttgcaatt cgcgcttttt caatattatt
     6661 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6721 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     6781 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcactcat
     6841 taggcacccc aggctttaca ctttatgct
//
