LOCUS       sequence-304678-        8846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8846)
  AUTHORS   Edie S, Zaghloul NA, Leitch CC, Klinedinst DK, Lebron J, Thole JF, 
            McCallion AS, Katsanis N, Reeves RH
  TITLE     Survey of Human Chromosome 21 Gene Expression Effects on Early 
            Development in Danio rerio.
  JOURNAL   G3 (Bethesda). 2018 Jul 2;8(7):2215-2223. doi: 
            10.1534/g3.118.200144.
  PUBMED    29760202
REFERENCE   2  (bases 1 to 8846)
  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: "10.1534/g3.118"; 
            journalName: "G3 (Bethesda)"; date: "2018-07-2- 2"; volume: "8"; 
            issue: "7"; pages: "2215-2223"
COMMENT     Sequence Label: TRPM2_pCSdest
FEATURES             Location/Qualifiers
     source          1..8846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1085..1109
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      5129..5138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5808..5832)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5876..6010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5900..5919)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5954..5973
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6123..6143)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6123..6141)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6162..6178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6162..6178)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6175..6197)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6186..6202
                     /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(6210..6216,6217..6234,6235..6240))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6255..6276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6393..6410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6564..7152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6644..6663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7323..8114,8115..8183))
                     /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     7946..7965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8184..8288)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8314..8769)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8451..8472)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8663..8682
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt cgaattcaag
     1081 gatcacaagt ttgtacaaaa aagcaggctc cgcggccgcc cccttcacca tggagccctc
     1141 agccctgagg aaagctggct cggagcagga ggagggcttt gaggggctgc ccagaagggt
     1201 cactgacctg gggatggtct ccaatctccg gcgcagcaac agcagcctct tcaagagctg
     1261 gaggctacag tgccccttcg gcaacaatga caagcaagaa agcctcagtt cgtggattcc
     1321 tgaaaacatc aagaagaaag aatgcgtgta ttttgtggaa agttccaaac tgtctgatgc
     1381 tgggaaggtg gtgtgtcagt gtggctacac gcatgagcag cacttggagg aggctaccaa
     1441 gccccacacc ttccagggca cacagtggga cccaaagaaa catgtccagg agatgccaac
     1501 cgatgccttt ggcgacatcg tcttcacggg cctgagccag aaggtgaaaa agtacgtccg
     1561 agtctcccag gacacgccct ccagcgtgat ctaccacctc atgacccagc actgggggct
     1621 ggacgtcccc aatctcttga tctcggtgac cgggggggcc aagaacttca acatgaagcc
     1681 gcggctgaag agcattttcc gcagaggcct ggtcaaggtg gctcagacca caggggcctg
     1741 gatcatcaca ggggggtccc acaccggcgt catgaagcag gtaggcgagg cggtgcggga
     1801 cttcagcctg agcagcagct acaaggaagg cgagctcatc accatcggag tcgccacctg
     1861 gggcactgtc caccgccgcg agggcctgat ccatcccacg ggcagcttcc ccgccgagta
     1921 catactggat gaggatggcc aagggaacct gacctgccta gacagcaacc actctcactt
     1981 catcctcgtg gacgacggga cccacggcca gtacggggtg gagattcctc tgaggaccag
     2041 gctggagaag ttcatatcgg agcagaccaa ggaaagagga ggtgtggcca tcaagatccc
     2101 catcgtgtgc gtggtgctgg agggcggccc gggcacgttg cacaccatcg acaacgccac
     2161 caccaacggc accccctgtg tggttgtgga gggctcgggc cgcgtggccg acgtcattgc
     2221 ccaggtggcc aacctgcctg tctcggacat cactatctcc ctgatccagc agaaactgag
     2281 cgtgttcttc caggagatgt ttgagacctt cacggaaagc aggattgtcg agtggaccaa
     2341 aaagatccaa gatatcgtcc ggaggcggca gctgctgact gtcttccggg aaggcaagga
     2401 tggtcagcag gacgtggatg tggccatctt gcaggccttg ctgaaagcct cacggagcca
     2461 agaccacttt ggccacgaga actgggacca ccagctgaaa ctggcagtgg catggaatcg
     2521 cgtggacatt gcccgcagtg agatcttcat ggatgagtgg cagtggaagc cttcagatct
     2581 gcaccccacg atgacagctg cactcatctc caacaagcct gagtttgtga agctcttcct
     2641 ggagaacggg gtgcagctga aggagtttgt cacctgggac accttgctct acctgtacga
     2701 gaacctggac ccctcctgcc tgttccacag caagctgcag aaggtgctgg tggaggatcc
     2761 cgagcgcccg gcttgcgcgc ccgcggcgcc ccgcctgcag atgcaccacg tggcccaggt
     2821 gctgcgggag ctgctggggg acttcacgca gccgctttat ccccggcccc ggcacaacga
     2881 ccggctgcgg ctcctgctgc ccgttcccca cgtcaagctc aacgtgcagg gagtgagcct
     2941 ccggtccctc tacaagcgtt cctcaggcca tgtgaccttc accatggacc ccatccgtga
     3001 ccttctcatt tgggccattg tccagaaccg tcgggagctg gcaggaatca tctgggctca
     3061 gagccaggac tgcatcgcag cggccttggc ctgcagcaag atcctgaagg aactgtccaa
     3121 ggaggaggag gacacggaca gctcggagga gatgctggcg ctggcggagg agtatgagca
     3181 cagagccatc ggggtcttca ccgagtgcta ccggaaggac gaagagagag cccagaaact
     3241 gctcacccgc gtgtccgagg cctgggggaa gaccacctgc ctgcagctcg ccctggaggc
     3301 caaggacatg aagtttgtgt ctcacggggg catccaggcc ttcctgacca aggtgtggtg
     3361 gggccagctc tccgtggaca atgggctgtg gcgtgtgacc ctgtgcatgc tggccttccc
     3421 gctgctcctc accggcctca tctccttcag ggagaagagg ctgcaggatg tgggcacccc
     3481 cgcggcccgc gcccgtgcct tcttcaccgc acccgtggtg gtcttccacc tgaacatcct
     3541 ctcctacttc gccttcctct gcctgttcgc ctacgtgctc atggtggact tccagcctgt
     3601 gccctcctgg tgcgagtgtg ccatctacct ctggctcttc tccttggtgt gcgaggagat
     3661 gcggcagctc ttctatgacc ctgacgagtg cgggctgatg aagaaggcag ccttgtactt
     3721 cagtgacttc tggaataagc tggacgtcgg cgcaatcttg ctcttcgtgg cagggctgac
     3781 ctgcaggctc atcccggcga cgctgtaccc cgggcgcgtc atcctctctc tggacttcat
     3841 cctgttctgc ctccggctca tgcacatttt taccatcagt aagacgctgg ggcccaagat
     3901 catcattgtg aagcggatga tgaaggacgt cttcttcttc ctcttcctgc tggctgtgtg
     3961 ggtggtgtcc ttcggggtgg ccaagcaggc catcctcatc cacaacgagc gccgggtgga
     4021 ctggctgttc cgaggggccg tctaccactc ctacctcacc atcttcgggc agatcccggg
     4081 ctacatcgac ggtgtgaact tcaacccgga gcactgcagc cccaatggca ccgaccccta
     4141 caagcctaag tgccccgaga gcgacgcgac gcagcagagg ccggccttcc ctgagtggct
     4201 gacggtcctc ctactctgcc tctacctgct cttcaccaac atcctgctgc tcaacctcct
     4261 catcgccatg ttcaactaca ccttccagca ggtgcaggag cacacggacc agatttggaa
     4321 gttccagcgc catgacctga tcgaggagta ccacggccgc cccgccgcgc cgcccccctt
     4381 catcctcctc agccacctgc agctcttcat caagagggtg gtcctgaaga ctccggccaa
     4441 gaggcacaag cagctcaaga acaagctgga gaagaacgag gaggcggccc tgctatcctg
     4501 ggagatctac ctgaaggaga actacctcca gaaccgacag ttccagcaaa agcagcggcc
     4561 cgagcagaag atcgaggaca tcagcaataa ggcagggctg gagctttggg ggtcaaggac
     4621 aagctctgtc tgcaaatcta gacagttcct gcatctcacg gtggtggaat cacgcgggtc
     4681 cgagaaggct ccagtaacca ccctcgcatg tttgcaaggc tgtctgggaa aacatggcag
     4741 ggttgacgcc atggtggacc tgctggacct ggacccactg aagaggtcgg gctccatgga
     4801 gcagaggttg gcctccctgg aggagcaggt ggcccagaca gcccgagccc tgcactggat
     4861 cgtgaggacg ctgcgggcca gcggcttcag ctcggaggcg gacgtcccca ctctggcctc
     4921 ccagaaggcc gcggaggagc cggatgctga gccgggaggc aggaagaaga cggaggagcc
     4981 gggcgacagc taccacgtga atgcccggca cctcctctac cccaactgcc ctgtcacgcg
     5041 cttccccgtg cccaacgaga aggtgccctg ggagacggag ttcctgatct atgacccacc
     5101 cttttacacg gcagagagga aggacgcggc cgccatggac cccatgggag acaccctgga
     5161 gccactgtcc acgatccagt acaacgtggt ggatggcctg agggaccgcc ggagcttcca
     5221 cgggccgtac acagtgcagg ccgggttgcc cctgaacccc atgggccgca caggactgcg
     5281 tgggcgcggg agcctcagct gcttcggacc caaccacacg ctgtacccca tggtcacgcg
     5341 gtggaggcgg aacgaggatg gagccatctg caggaagagc ataaagaaga tgctggaagt
     5401 gctggtggtg aagctccctc tctccgagca ctgggccctg cctgggggct cccgggagcc
     5461 aggggagatg ctacctcgga agctgaagcg gatcctccgg caggagcact ggccgtcttt
     5521 tgaaaacttg ctgaagtgcg gcatggaggt gtacaaaggc tacatggatg acccgaggaa
     5581 cacggacaat gcctggatcg agacggtggc cgtcagcgtc cacttccagg accagaatga
     5641 cgtggagctg aacaggctga actctaacct gcacgcctgc gactcggggg cctccatccg
     5701 atggcaggtg gtggacaggc gcatcccact ctatgcgaac cacaagaccc tcctccagaa
     5761 ggcagccgct gagttcgggg ctcactactg aaagggtggg cgcgccgacc cagctttctt
     5821 gtacaaagtg gtgatcctct cgagcctcta gaactatagt gagtcgtatt acgtagatcc
     5881 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5941 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     6001 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     6061 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     6121 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     6181 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     6241 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6301 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6361 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6421 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6481 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6541 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6601 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6661 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6721 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6781 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6841 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6901 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6961 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7021 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7081 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7141 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7201 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7261 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7321 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7381 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7441 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7501 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7561 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7621 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7681 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7741 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7801 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7861 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7921 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7981 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8041 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8101 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8161 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8221 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8281 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     8341 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     8401 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     8461 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     8521 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     8581 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     8641 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     8701 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     8761 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     8821 cctcttcgct attacgccag tcgacc
//
