LOCUS       sequence-468736-        6139 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 6139)
  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 6139)
  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: MX1_pCSdest
FEATURES             Location/Qualifiers
     source          1..6139
                     /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(2124..2148)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2192..2326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2216..2235)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2270..2289
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2439..2459)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2439..2457)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2478..2494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2478..2494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2491..2513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2502..2518
                     /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(2526..2532,2533..2550,2551..2556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2571..2592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2709..2726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2880..3467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2960..2979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3638..4429,4430..4498))
                     /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     4261..4280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4499..4603)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4629..5084)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4766..4787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4978..4997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5155..6139
                     /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 accatggttg tttccgaagt ggacatcgca aaagctgatc cagctgctgc
      181 atcccaccct ctattactga atggagatgc tactgtggcc cagaaaaatc caggctcggt
      241 ggctgagaac aacctgtgca gccagtatga ggagaaggtg cgcccctgca tcgacctcat
      301 tgactccctg cgggctctag gtgtggagca ggacctggcc ctgccagcca tcgccgtcat
      361 cggggaccag agctcgggca agagctccgt gttggaggca ctgtcaggag ttgcccttcc
      421 cagaggcagc gggatcgtga ccagatgccc gctggtgctg aaactgaaga aacttgtgaa
      481 cgaagataag tggagaggca aggtcagtta ccaggactac gagattgaga tttcggatgc
      541 ttcagaggta gaaaaggaaa ttaataaagc ccagaatgcc atcgccgggg aaggaatggg
      601 aatcagtcat gagctaatca ccctggagat cagctcccga gatgtcccgg atctgactct
      661 aatagacctt cctggcataa ccagagtggc tgtgggcaat cagcctgctg acattgggta
      721 taagatcaag acactcatca agaagtacat ccagaggcag gagacaatca gcctggtggt
      781 ggtccccagt aatgtggaca ttgccaccac agaggctctc agcatggccc aggaggtgga
      841 ccccgaggga gacaggacca tcggaatctt gacgaagcct gatctggtgg acaaaggaac
      901 tgaagacaag gttgtggacg tggtgcggaa cctcgtgttc cacctgaaga agggttacat
      961 gattgtcaag tgccggggcc agcaggagat ccaggaccag ctgagcctgt ccgaagccct
     1021 gcagagagag aagatcttct ttgagaacca cccatatttc agggatctgc tggaggaagg
     1081 aaaggccacg gttccctgcc tggcagaaaa acttaccagc gagctcatca cacatatctg
     1141 taaatctctg cccctgttag aaaatcaaat caaggagact caccagagaa taacagagga
     1201 gctacaaaag tatggtgtcg acataccgga agacgaaaat gaaaaaatgt tcttcctgat
     1261 agataaaatt aatgccttta atcaggacat cactgctctc atgcaaggag aggaaactgt
     1321 aggggaggaa gacattcggc tgtttaccag actccgacac gagttccaca aatggagtac
     1381 aataattgaa aacaattttc aagaaggcca taaaattttg agtagaaaaa tccagaaatt
     1441 tgaaaatcag tatcgtggta gagagctgcc aggctttgtg aattacagga catttgagac
     1501 aatcgtgaaa cagcaaatca aggcactgga agagccggct gtggatatgc tacacaccgt
     1561 gacggatatg gtccggcttg ctttcacaga tgtttcgata aaaaattttg aagagttttt
     1621 taacctccac agaaccgcca agtccaaaat tgaagacatt agagcagaac aagagagaga
     1681 aggtgagaag ctgatccgcc tccacttcca gatggaacag attgtctact gccaggacca
     1741 ggtatacagg ggtgcattgc agaaggtcag agagaaggag ctggaagaag aaaagaagaa
     1801 gaaatcctgg gattttgggg ctttccaatc cagctcggca acagactctt ccatggagga
     1861 gatctttcag cacctgatgg cctatcacca ggaggccagc aagcgcatct ccagccacat
     1921 ccctttgatc atccagttct tcatgctcca gacgtacggc cagcagcttc agaaggccat
     1981 gctgcagctc ctgcaggaca aggacaccta cagctggctc ctgaaggagc ggagcgacac
     2041 cagcgacaag cggaagttcc tgaaggagcg gcttgcacgg ctgacgcagg ctcggcgccg
     2101 gcttgcccag ttccccggtt aaaacccagc tttcttgtac aaagtggtga tcctctcgag
     2161 cctctagaac tatagtgagt cgtattacgt agatccagac atgataagat acattgatga
     2221 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2281 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2341 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc
     2401 ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     2461 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2521 ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2581 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2641 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2701 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2761 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2821 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2881 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2941 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3001 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3061 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3121 aagctgggct gtgtgcacga acccccgttc agcccgaccg ctgcgcctta tccggtaact
     3181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4141 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4201 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4261 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4321 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4381 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4441 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4501 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4561 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4621 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4681 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4741 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4801 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     4861 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4921 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4981 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     5041 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     5101 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac
     5161 cgccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg
     5221 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc
     5281 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg
     5341 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct
     5401 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata
     5461 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc
     5521 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat
     5581 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata
     5641 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga
     5701 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt
     5761 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca
     5821 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg
     5881 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta
     5941 ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat
     6001 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca
     6061 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag
     6121 caatgctcgt ttagggaac
//
