LOCUS       sequence-155809-        4444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains aTc inducible Scaffold-mCherry-AAV and TetR genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4444)
  AUTHORS   Hsiao V, de los Santos EL, Whitaker WR, Dueber JE, Murray RM
  TITLE     Design and implementation of a biomolecular concentration tracker.
  JOURNAL   ACS Synth Biol. 2015 Feb 20;4(2):150-61. doi: 10.1021/sb500024b. 
            Epub 2014 May 15.
  PUBMED    24847683
REFERENCE   2  (bases 1 to 4444)
  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"; doi: "10.1021/sb500024b";
            journalName: "ACS Synth Biol"; date: "2015-02-20- 20"; volume: "4"; 
            issue: "2"; pages: "150-61"
COMMENT     Sequence Label: pVH001
FEATURES             Location/Qualifiers
     source          1..4444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..83
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    64..82
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             241..258
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             511..1218
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(656..674)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     924..943
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1286..1315
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1331..1348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1404..1421)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1574..1620
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1662..1679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1833..2421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1913..1932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2592..3383,3384..3452))
                     /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     3215..3234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3453..3557)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(3647..3690)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(join(3830..4444,1..9))
                     /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     4361..4380
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 tcttgacatc attaattcct aatttttgtt gacactctat cgttgataga gttattttac
       61 cactccctat cagtgataga gaaaagtgaa aatccatatg actagcagat ccactagagt
      121 cgactaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      181 ctttcgtctt cacgctagcg gtacctgaag tgtggcccaa cgcaaactaa gagatctatg
      241 catcaccatc accatcacga ctacgacatc ccgactaccg aaaacctgta cttccagcct
      301 ccacctgcat taccgccaaa gagaaggcgt ggatctgggt ccggcagcgg tggatctatc
      361 acaattcggg cagcttttct ggagaaggaa aacaccgcac tgcgtactga gattgctgaa
      421 ctggagaaag aagtaggccg ctgcgagaat attgttagca aatatgaaac ccgctacggc
      481 cccctgggat ccggtagtgg cagcggcagc atggtgagca agggcgagga ggataacatg
      541 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
      601 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
      661 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
      721 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
      781 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
      841 accgtgaccc aggactcctc cttgcaggac ggcgagttca tctacaaggt gaagctgcgc
      901 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
      961 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     1021 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     1081 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     1141 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     1201 atggacgagc tgtacaaggc agcaaacgac gaaaactacg ctgcagcagt ttaataataa
     1261 ctcgagtaat gagaagcttg ggcccgaaca aaaactcatc tcagaagagg atctgaatag
     1321 cgccgtcgac catcatcatc atcatcattg agtttaaacg gtctccagct tggctgtttt
     1381 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     1441 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     1501 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     1561 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     1621 ctgttgtttg tcggtgaact aatttctaga ctgcaggctt cctcgctcac tgactcgctg
     1681 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1741 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1921 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2221 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     2281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2401 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2461 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2521 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2581 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2641 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2701 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2761 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2821 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2881 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2941 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3001 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3061 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3121 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3181 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3241 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3301 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3361 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3421 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3481 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3541 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3601 attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa
     3661 aaatccttag ctttcgctaa ggatgatttc tggaattcac tagtgatccg tttccattta
     3721 ggtgggtacg ttggagccgc attattttcg ctttatgaat ctaaagggtg gttaactcga
     3781 catcttggtt accgtgaagt taccatcacg gaaaaaggtt atgctgcttt taagacccac
     3841 tttcacattt aagttgtttt tctaatccgc atatgatcaa ttcaaggccg aataagaagg
     3901 ctggctctgc accttggtga tcaaataatt cgatagcttg tcgtaataat ggcggcatac
     3961 tatcagtagt aggtgtttcc ctttcttctt tagcgacttg atgctcttga tcttccaata
     4021 cgcaacctaa agtaaaatgc cccacagcgc tgagtgcata taatgcattc tctagtgaaa
     4081 aaccttgttg gcataaaaag gctaattgat tttcgagagt ttcatactgt ttttctgtag
     4141 gccgtgtacc taaatgtact tttgctccat cgcgatgact tagtaaagca catctaaaac
     4201 ttttagcgtt attacgtaaa aaatcttgcc agctttcccc ttctaaaggg caaaagtgag
     4261 tatggtgcct atctaacatc tcaatggcta aggcgtcgag caaagcccgc ttatttttta
     4321 catgccaata caatgtaggc tgctctacac ctagcttctg ggcgagttta cgggttgtta
     4381 aaccttcgat tccgacctca ttaagcagct ctaatgcgct gttaatcact ttacttttat
     4441 ctaa
//
