LOCUS       sequence-314862-        6479 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 6479)
  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 6479)
  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: DYRK1A_pCSdest
FEATURES             Location/Qualifiers
     source          1..6479
                     /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"
     CDS             2948..2977
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2951..2980
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2954..2983
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2957..2986
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     protein_bind    complement(3441..3465)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3509..3643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3533..3552)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3587..3606
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3756..3776)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3756..3774)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3795..3811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3795..3811)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3808..3830)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3819..3835
                     /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(3843..3849,3850..3867,3868..3873))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3888..3909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4026..4043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4197..4785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4277..4296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4956..5747,5748..5816))
                     /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     5579..5598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5817..5921)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5947..6402)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6084..6105)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6296..6315
                     /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 tgcatacagg
     1141 aggagagact tcagcatgca aaccttcatc tgttcggctt gcaccgtcat tttcattcca
     1201 tgctgctggc cttcagatgg ctggacagat gccccattca catcagtaca gtgaccgtcg
     1261 ccagccaaac ataagtgacc aacaggtttc tgccttatca tattctgacc agattcagca
     1321 acctctaact aaccaggtga tgcctgatat tgtcatgtta cagaggcgga tgccccaaac
     1381 cttccgtgac ccagcaactg ctcccctgag aaaactttct gttgacttga tcaaaacata
     1441 caagcatatt aatgaggttt actatgcaaa aaagaagcga agacaccaac agggccaggg
     1501 agacgattct agtcataaga aggaacggaa ggtttacaat gatggttatg atgatgataa
     1561 ctatgattat attgtaaaaa acggagaaaa gtggatggat cgttacgaaa ttgactcctt
     1621 gataggcaaa ggttcctttg gacaggttgt aaaggcatat gatcgtgtgg agcaagaatg
     1681 ggttgccatt aaaataataa agaacaagaa ggcttttctg aatcaagcac agatagaagt
     1741 gcgacttctt gagctcatga acaaacatga cactgaaatg aaatactaca tagtgcattt
     1801 gaaacgccac tttatgtttc gaaaccatct ctgtttagtt tttgaaatgc tgtcctacaa
     1861 cctctatgac ttgctgagaa acaccaattt ccgaggggtc tctttgaacc taacacgaaa
     1921 gtttgcgcaa cagatgtgca ctgcactgct tttccttgcg actccagaac ttagtatcat
     1981 tcactgtgat ctaaaacctg aaaatatcct tctttgtaac cccaaacgca gtgcaatcaa
     2041 gatagttgac tttggcagtt cttgtcagtt ggggcagagg atataccagt atattcagag
     2101 tcgcttttat cggtctccag aggtgctact gggaatgcct tatgaccttg ccattgatat
     2161 gtggtccctc gggtgtattt tggttgaaat gcacactgga gaacctctgt tcagtggtgc
     2221 caatgaggta gatcagatga ataaaatagt ggaagttctg ggtattccac ctgctcatat
     2281 tcttgaccaa gcaccaaaag caagaaagtt ctttgagaag ttgccagatg gcacttggaa
     2341 cttaaagaag accaaagatg gaaaacggga gtacaaacca ccaggaaccc gtaaacttca
     2401 taacattctt ggagtggaaa caggaggacc tggtgggcga cgtgctgggg agtcaggtca
     2461 tacggtcgct gactacttga agttcaaaga cctcatttta aggatgcttg attatgaccc
     2521 caaaactcga attcaacctt attatgctct gcagcacagt ttcttcaaga aaacagctga
     2581 tgaaggtaca aatacaagta atagtgtatc tacaagcccc gccatggagc agtctcagtc
     2641 ttcgggcacc acctccagta catcgtcaag ctcaggtggc tcatcgggga caagcaacag
     2701 tgggagagcc cggtcggatc cgacgcacca gcatcggcac agtggtgggc acttcacagc
     2761 tgccgtgcag gccatggact gcgagacaca cagtccccag gtgcgtcagc aatttcctgc
     2821 tcctcttggt tggtcaggca ctgaagctcc tacacaggtc actgttgaaa ctcatcctgt
     2881 tcaagaaaca acctttcatg tagcccctca acagaatgca ttgcatcatc accatggtaa
     2941 cagttcccat caccatcacc accaccacca ccatcaccac caccatggac aacaagcctt
     3001 gggtaaccgg accaggccaa gggtctacaa ttctccaacg aatagctcct ctacccaaga
     3061 ttctatggag gttggccaca gtcaccactc catgacatcc ctgtcttcct caacgacttc
     3121 ttcctcgaca tcttcctcct ctactggtaa ccaaggcaat caggcctacc agaatcgccc
     3181 agtggctgct aataccttgg actttggaca gaatggagct atggacgtta atttgaccgt
     3241 ctactccaat ccccgccaag agactggcat agctggacat ccaacatacc aattttctgc
     3301 taatacaggt cctgcacatt acatgactga aggacatctg acaatgaggc aaggggctga
     3361 tagagaagag tcccccatga caggagtttg tgtgcaacag agtcctgtag ctagctcgta
     3421 atagaagggt gggcgcgccg acccagcttt cttgtacaaa gtggtgatcc tctcgagcct
     3481 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     3541 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3601 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3661 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     3721 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     3781 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3841 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3901 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3961 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4201 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4621 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5821 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5881 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5941 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     6001 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     6061 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     6121 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     6181 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6241 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6301 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6361 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     6421 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacc
//
