LOCUS       sequence-340014-        7178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian acoustic reporter gene 1 cassette 1 (expressing GvpB).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7178)
  AUTHORS   Howells AR, Welch PJ, Kim J, Forest CR, Shi C, Lian XL
  TITLE     A drug-selectable acoustic reporter gene system for human cell 
            ultrasound imaging.
  JOURNAL   Bioeng Transl Med. 2023 Aug 2;9(2):e10584. doi: 10.1002/btm2.10584. 
            eCollection 2024 Mar.
  PUBMED    38435822
REFERENCE   2  (bases 1 to 7178)
  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: "Bioeng 
            Transl Med."; date: "2023-08-2"; volume: "9(2):e10584. doi"; pages: 
            " 10.1002/btm2.10584. eCollection 2024 Mar"
COMMENT     Sequence Label: XLone-GvpB
FEATURES             Location/Qualifiers
     source          1..7178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..395
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    36..54
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    72..90
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    108..126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    144..162
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    180..198
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    216..234
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    252..270
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    705..1256
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(849..866)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1076..1095
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1268..1978
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVNTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1413..1431)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1681..1700
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2159..2293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2569..2603
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2614..2631)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2614..2630)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2623..2645)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2839..2858)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2958..2980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3018..3036)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3104..3208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3209..3277,3278..4069)
                     /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     complement(3427..3446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4240..4828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4729..4748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4982..4999
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5116..5137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5152..5157,5158..5175,5176..5182)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5190..5206
                     /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     5195..5217
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5214..5230
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5214..5230
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5256..5318
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    5494..5575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5692..6090)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(6091..6147)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(6157..6900)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             complement(6901..6924)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      6924..6933
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6953..7164)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 gagccactcg agtggaatta tcacctcgag tttactccct atcagtgata gagaacgtat
       61 gaagagttta ctccctatca gtgatagaga acgtatgcag actttactcc ctatcagtga
      121 tagagaacgt ataaggagtt tactccctat cagtgataga gaacgtatga ccagtttact
      181 ccctatcagt gatagagaac gtatctacag tttactccct atcagtgata gagaacgtat
      241 atccagttta ctccctatca gtgatagaga acgtataagc tttgcttatg taaaccaggg
      301 cgcctataaa agagtgctga ttttttgagt aaacttcaat tccacaacac ttttgtctta
      361 taccaacttt ccgtaccact tcctaccctc gtaaaggtac cgccaccatg agcatccaga
      421 agtccaccaa cagcagcagc ctggccgaag tgatcgaccg gatcctggac aagggcatcg
      481 tgatcgacgc cttcgccaga gtgtccgtcg tgggcatcga gatcctgacc atcgaggcca
      541 gagtcgtgat cgccagcgtg gacacctggc tgagatatgc cgaagccgtg ggcctgctgc
      601 gggacgacgt ggaagaaaat ggcctgcccg agcggagcaa cagctctgag ggacagcccc
      661 ggttcagcat ctgaactaaa tcgcactgtc ggcgtccccc cctaacgtta ctggccgaag
      721 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
      781 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
      841 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
      901 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
      961 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     1021 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     1081 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     1141 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaacgt
     1201 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     1261 cacaaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     1321 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1381 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
     1441 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     1501 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     1561 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     1621 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     1681 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     1741 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     1801 cgacgctgag gtcaatacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     1861 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     1921 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtgaac
     1981 tagtagacca cctcccctgc gagctaagct ggacagccaa tgacgggtaa gagagtgaca
     2041 tttttcacta acctaagaca ggagggccgt cagagctact gcctaatcca aagacgggta
     2101 aaagtgataa aaatgtatca ctccaaccta agacaggcgc agcttccgag ggatttgaga
     2161 tccagacatg ataagataca ttgatgagtt tggacaaacc aaaactagaa tgcagtgaaa
     2221 aaaatgcctt atttgtgaaa tttgtgatgc tattgcctta tttgtaacca ttataagctg
     2281 caataaacaa gtttgatatc tataacaaga aaatatatat ataataagtt atcacgtaag
     2341 tagaacatga aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag
     2401 ataatcatgc gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc
     2461 gcattgacaa gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca
     2521 gcgacggatt cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt
     2581 ttacgcagac tatctttcta gggttaagaa ttcactggcc gtcgttttac aacgtcgtga
     2641 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2701 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2761 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2821 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     2881 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     2941 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3001 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3061 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3421 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3721 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4381 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4981 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5041 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5101 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5161 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5221 cagctatgac catgattacg ccaaggtcga cttaacccta gaaagataat catattgtga
     5281 cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg
     5341 aggtttattt attaatttga atagatatta agttttatta tatttacact tacatactaa
     5401 taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa aacaaaaact
     5461 caaaatttct tctataaagt aacaaaactt ttagcagtga aaaaaatgct ttatttgtga
     5521 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     5581 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     5641 caagtaaaac ctctacaaat gtggtatggc tgattatgat cctctggaga tttagccctc
     5701 ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct gtccatcact
     5761 gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc
     5821 gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag tcaggttgcc
     5881 agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc ccccagtaaa
     5941 atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg cgctggcgac
     6001 gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt caatgagggt
     6061 ggattcttct tgagacaaag gcttggccat ggggccgggg ttctcctcca cgtcgccggc
     6121 ctgcttcagc aggctgaagt tggtggcgcc gctgcccccg gggagcatgt caaggtcaaa
     6181 atcgtcaaga gcgtcagcag gcagcatatc aaggtcaaag tcgtcaaggg catcggctgg
     6241 gagcatgtct aagtcaaaat cgtcaagggc gtcggtcggc ccgccgcttt cgcactttag
     6301 ctgtttctcc aggccacata tgattagttc caggccgaaa aggaaggcag gttcggctcc
     6361 ctgccggtcg aacagctcaa ttgcttgttt cagaagtggg ggcatagaat cggtggtagg
     6421 tgtctctctt tcctcttttg ctacttgatg ctcctgttcc tccaatacgc agcccagtgt
     6481 aaagtggccc acggcggaca gagcgtacag tgcgttctcc agggagaagc cttgctgaca
     6541 caggaacgcg agctgatttt ccagggtttc gtactgtttc tctgttgggc gggtgccgag
     6601 atgcacttta gccccgtcgc gatgtgagag gagagcacag cggtatgact tggcgttgtt
     6661 ccgcagaaag tcttgccatg actcgccttc cagggggcag gagtgggtat gatgcctgtc
     6721 cagcatctcg attggcaggg catcgagcag ggcccgcttg ttcttcacgt gccagtacag
     6781 ggtaggctgc tcaactccca gcttttgagc gagtttcctt gtcgtcaggc cttcgatacc
     6841 gactccattg agtaattcca gagcagagtt tatgactttg ctcttgtcca gtctagacat
     6901 cttatcgtca tcgtctttgt aatccatggt ggcggatccc gcgtcacgac acctgtgttc
     6961 tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt atatacggtt
     7021 ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc cagtttaccc
     7081 cgcgccacct tctctaggca ccggttcaat tgccgacccc tccccccaac ttctcgggga
     7141 ctgtgggcga tgtgcgctct gcccactgac gggcaccg
//
