LOCUS       sequence-473205-        6674 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 6674)
  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 6674)
  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: RIPK4_pCSdest
FEATURES             Location/Qualifiers
     source          1..6674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    107..131
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2658..2682)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2726..2860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2750..2769)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2804..2823
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2973..2993)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2973..2991)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3012..3028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3012..3028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3025..3047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3036..3052
                     /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(3060..3066,3067..3084,3085..3090))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3105..3126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3243..3260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3414..4002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3494..3513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4173..4964,4965..5033))
                     /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     4796..4815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5034..5138)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5164..5619)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5301..5322)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5513..5532
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5690..6674
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggatcacaa gtttgtacaa
      121 aaaagcaggc tttaaaggaa ccatctgcag aattcgccct tggcgctggc tagacgtgcg
      181 cgatggaggg cgacggcggg accccatggg ccctggcgct gctgcgcacc ttcgacgcgg
      241 gcgagttcac gggctgggag aaggtgggct cgggcggctt cgggcaggtg tacaaggtac
      301 gccatgtcca ctggaagacc tggctggcca tcaagtgctc gcccagcctg cacgtcgacg
      361 acagggagcg catggagctt ttggaagaag ccaagaagat ggagatggcc aagtttcgct
      421 acatcctgcc tgtgtatggc atctgccgcg aacctgtcgg cctggtcatg gagtacatgg
      481 agacgggctc cctggaaaag ctgctggctt cggagccatt gccatgggat ctccggttcc
      541 gaatcatcca cgagacggcg gtgggcatga acttcctgca ctgcatggcc ccgccactcc
      601 tgcacctgga cctcaagccc gcgaacatcc tgctggatgc ccactaccac gtcaagattt
      661 ctgattttgg tctggccaag tgcaacgggc tgtcccactc gcatgacctc agcatggatg
      721 gcctgtttgg cacaatcgcc tacctccctc cagagcgcat cagggagaag agccggctct
      781 tcgacaccaa gcacgatgta tacagctttg cgatcgtcat ctggggcgtg ctcacacaga
      841 agaagccgtt tgcagatgag aagaacatcc tgcacatcat ggtgaaggtg gtgaagggcc
      901 accgccccga gctgccgccc gtgtgcagag cccggccgcg cgcctgcagc cacctgatac
      961 gcctcatgca gcggtgctgg cagggggatc cgcgagttag gcccaccttc caagaaatta
     1021 cttctgaaac cgaggacctg tgtgaaaagc ctgatgacga agtgaaagaa actgctcatg
     1081 atctggacgt gaaaagcccc ccggagccca ggagcgaggt ggtgcctgcg aggctcaagc
     1141 gggcctctgc ccccaccttc gataacgact acagcctctc cgagctgctc tcacagctgg
     1201 actctggagt ttcccaggct gtcgagggcc ccgaggagct cagccgcagc tcctctgagt
     1261 ccaagctgcc atcgtccggc agtgggaaga ggctctcggg ggtgtcctcg gtggactccg
     1321 ccttctcttc cagaggatca ctgtcgctgt cctttgagcg ggaaccttca accagcgatc
     1381 tgggcaccac agacgtccag aagaagaagc ttgtggatgc catcgtgtcc ggggacacca
     1441 gcaaactgat gaagatcctg cagccgcagg acgtggacct ggcactggac agcggtgcca
     1501 gcctgctgca cctggcggtg gaggccgggc aagaggagtg cgccaagtgg ctgctgctca
     1561 acaatgccaa ccccaacctg agcaaccgta ggggctccac cccgttgcac atggccgtgg
     1621 agaggagggt gcggggtgtc gtggagctcc tgctggcgcg gaagatcagt gtcaacgcca
     1681 aggatgagga ccagtggaca gccctccact ttgcagccca gaacggggac gagtctagca
     1741 cacggctgct gttggagaag aacgcctcgg tcaacgaggt ggactttgag ggccggacgc
     1801 ccatgcacgt ggcctgccag cacgggcagg agaatatcgt gcgcatcctg ctgcgccgag
     1861 gcgtggacgt gagcctgcag ggcaaggatg cctggctgcc actgcactac gctgcctggc
     1921 agggccacct gcccatcgtc aagctgctgg ccaagcagcc gggggtgagt gtgaacgccc
     1981 agacgctgga tgggaggacg ccattgcacc tggccgcaca gcgcgggcac taccgcgtgg
     2041 cccgcatcct catcgacctg tgctccgacg tcaacgtctg cagcctgctg gcacagacac
     2101 ccctgcacgt ggccgcggag acggggcaca cgagcactgc caggctgctc ctgcatcggg
     2161 gcgctggcaa ggaggccgtg acctcagacg gctacaccgc tctgcacctg gctgcccgca
     2221 acggacacct ggccactgtc aagctgcttg tcgaggagaa ggccgatgtg ctggcccggg
     2281 gacccctgaa ccagacggcg ctgcacctgg ctgccgccca cgggcactcg gaggtggtgg
     2341 aggagttggt cagcgccgat gtcattgacc tgttcgacga gcaggggctc agcgcgctgc
     2401 acctggccgc ccagggccgg cacgcacaga cggtggagac tctgctcagg catggggccc
     2461 acatcaacct gcagagcctc aagttccagg gcggccatgg ccccgccgcc acactcctgc
     2521 ggcgaagcaa gacctagctg gctgcctgcg gagaccgggg gtccacgtgg ggctcttgtc
     2581 ctgtcctgtg ttcctcgtgg ggatggaacg atcaatggtt gaagggcgaa ttccagcaca
     2641 ctggcggccg ttactagacc cagctttctt gtacaaagtg gtgatcctct cgagcctcta
     2701 gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg
     2761 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2821 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2881 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc
     2941 aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     3001 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3061 aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3241 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3301 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3421 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3481 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3541 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3601 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3661 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3721 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3781 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3841 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3901 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3961 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4021 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4081 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4141 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4201 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4261 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4321 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4381 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4441 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4501 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4561 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4621 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4681 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4741 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4801 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4861 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4921 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4981 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5041 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5101 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5161 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5221 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5281 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5341 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5401 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5461 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5521 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5581 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     5641 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca
     5701 attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca
     5761 attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca
     5821 attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca
     5881 actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc
     5941 aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca
     6001 ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt
     6061 caatactatg tattggccct atgccatata gtattccata tatgggtttt cctattgacg
     6121 tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc
     6181 catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca
     6241 tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg
     6301 gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg
     6361 atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa
     6421 tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt
     6481 ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc
     6541 gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg
     6601 gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg
     6661 ctcgtttagg gaac
//
