LOCUS       sequence-376580-        8142 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 8142)
  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 8142)
  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)
FEATURES             Location/Qualifiers
     source          1..8142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1162..1617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1511..1530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1744..1766
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1758..1775
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1759..1775
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1905..1967
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        2401..2750
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    2409..2427
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2445..2463
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2481..2499
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2517..2535
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2553..2571
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2589..2607
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2625..2643
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2689..2713
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2764..2773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3020..3571
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3164..3181)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3391..3410
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(4356..4375)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4357..4375)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4770..5358
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4823..4843)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5241..5252)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5378..5854
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5577..5596)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   6627..6661
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7152..7168)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7152..7168)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7165..7187)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7176..7192
                     /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)."
     promoter        complement(join(7200..7206,7207..7224,7225..7230))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7245..7266
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7383..7400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7554..8142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7634..7653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     1621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1681 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     1741 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcctcgttc attcacgttt
     1801 ttgaacccgt ggaggacggg cagactcgcg gtgcaaatgt gttttacagc gtgatggagc
     1861 agatgaagat gctcgacacg ctgcagaaca cgcagctaga ttaaccctag aaagataatc
     1921 atattgtgac gtacgttaaa gataatcatg tgtaaaattg acgcatgtgt tttatcggtc
     1981 tgtatatcga ggtttattta ttaatttgaa tagatattaa gttttattat atttacactt
     2041 acatactaat aataaattca acaaacaatt tatttatgtt tatttattta ttaaaaaaaa
     2101 caaaaactca aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc
     2161 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     2221 gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag
     2281 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     2341 gagcctgcag acacctgggg ggatacgggg aaaaggcctc cacggccact agtttcactc
     2401 gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag
     2461 agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc
     2521 tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta
     2581 cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag
     2641 agaacgtatg tcgaggtagg cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt
     2701 gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatactt ggtacctatg
     2761 tccgccacca tggccgtgga aaagaccaac agcagcagct ccctggccga agtgatcgac
     2821 agaatcctgg acaagggcat cgtgatcgac gcctgggtgc gcgtgtccct cgtgggaatt
     2881 gagctgctgg ccatcgaggc ccggatcgtg attgccagcg tggaaaccta cctgaagtac
     2941 gccgaggccg tgggcctgac acagagtgct gctgtgcctg cttgataata ctaaatcgca
     3001 ctgtcggcgt ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     3061 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     3121 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     3181 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     3241 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     3301 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     3361 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     3421 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     3481 tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg
     3541 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgtc caagggcgag
     3601 gaactgttca ccggcgtggt gcccatcctg gtggaactgg atggcgacgt gaacggccac
     3661 aagttctctg tgcggggaga gggcgaaggc gacgccacaa atggcaagct gaccctgaag
     3721 ttcatctgca ccaccggcaa gctgcccgtg ccttggccta ccctcgtgac cacactgtct
     3781 cacggcgtgc agtgcttcgc cagatacccc gaccacatga agcagcacga tttcttcaag
     3841 agcgccatgc ccgagggcta cgtgcaggaa cggaccatct tcttcaagga cgacggcacc
     3901 tacaagacca gagccgaagt gaagttcgag ggcgacaccc tcgtgaaccg gatcgagctg
     3961 aagggcgtgg acttcaaaga ggacggcaac atcctgggcc acaagctgga gtacaacttc
     4021 aacagccaca acatctacat catggccgtg aagcagaaaa acggcatcaa agtgaacttc
     4081 aagatccggc acaacgtgga agatggcagc gtgcagctgg ccgaccacta ccagcagaac
     4141 acccccatcg gagatggccc cgtgctgctg cctgatagcc actacctgag cacccagagc
     4201 gtgctgagca aggaccccaa cgagaagcgg gaccacatgg tgctgctgga atttcggacc
     4261 gccgctggca tcaccctggg catggatgag ctgtacaagt gaaggatccc tcgaggggcc
     4321 caagcttacg cgtgcatgcg acgtcatagc tctctcccta tagtgagtcg tattataagc
     4381 tagcttggga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt ggacaaacta
     4441 cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa
     4501 ctagctgcat atgcttgctg cttgagagtt ttgcttactg agtatgattt atgaaaatat
     4561 tatacacagg agctagtgat tctaattgtt tgtgtatttt agattcacag tcccaaggct
     4621 catttcaggc ccctcagtcc tcacagtctg ttcatgatca taatcagcca taccacattt
     4681 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4741 atgaatgcaa ttgttgttgt tcgtggctga atcaacctct ggattacaaa atttgtgaaa
     4801 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4861 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4921 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4981 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5041 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5101 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     5161 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5221 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5281 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     5341 tttgggccgc ctccccgcca gagacaatta acttcgcggg tggcatccct gtgacccctc
     5401 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     5461 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     5521 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     5581 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     5641 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     5701 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     5761 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     5821 acaggcgtga accactgctc ccttccctgt ccttgaagtt ctcatcatgt ctggaattga
     5881 ctcaaatgat gtcaattagt ctatcagaag ctcatctggt ctcccttccg ggggacaaga
     5941 catccctgtt taatatttaa acagcagtgt tcccaaactg ggttcttata tcccttgctc
     6001 tggtcaacca ggttgcaggg tttcctgtcc tcacaggaac gaagtcccta aagaaacagt
     6061 ggcagccagg tttagccccg gaattgactg gattcctttt ttagggccca ttggtatggc
     6121 tttttccccg tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag
     6181 gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg
     6241 ggatgcgggg ggagcgccgg accggagcgg agccccgggc ggctcgctgc tgccccctag
     6301 cgggggaggg acgtaattac atccctgggg gctttggggg ggggctgtcc ctgatatcta
     6361 taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata
     6421 attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact
     6481 cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg
     6541 ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag
     6601 aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg
     6661 gttaatctag ctgcatcagg atcatatcgt cgggtctttt ttccggctca gtcatcgccc
     6721 aagctggcgc tatctgggca tcggggagga agaagcccgt gccttttccc gcgaggttga
     6781 agcggcatgg aaagagtttg ccgaggatga ctgctgctgc attgacgttg agcgaaaacg
     6841 cacgtttacc atgatgattc gggaaggtgt ggccatgcac gcctttaacg gtgaactgtt
     6901 cgttcaggcc acctgggata ccagttcgtc gcggcttttc cggacacagt tccggatggt
     6961 cagcccgaag cgcatcagca acccgaacaa taccggcgac agccggaact gccgtgccgg
     7021 tgtgcagatt aatgacagcg gtgcggcgct gggatattac gtcagcgagg acgggtatcc
     7081 tggctggatg ccgcagaaat ggacatggat accccgtgag ttacccggcg ggcgcgcttg
     7141 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     7201 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     7261 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7321 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7381 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7441 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7501 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7561 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7621 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7681 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7741 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7801 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7861 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7921 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7981 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     8041 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     8101 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aa
//
