LOCUS       sequence-235411-        6299 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 6299)
  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 6299)
  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: MX2_pCSdest
FEATURES             Location/Qualifiers
     source          1..6299
                     /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"
     protein_bind    complement(3261..3285)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3329..3463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3353..3372)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3407..3426
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3576..3596)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3576..3594)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3615..3631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3615..3631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3628..3650)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3639..3655
                     /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(3663..3669,3670..3687,3688..3693))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3708..3729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3846..3863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4017..4605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4097..4116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4776..5567,5568..5636))
                     /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     5399..5418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5637..5741)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5767..6222)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5904..5925)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6116..6135
                     /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 aagcaggcac catgtctaag gcccacaagc cttggcccta
     1141 ccggaggaga agtcaatttt cttctcgaaa atacctgaaa aaagaaatga attccttcca
     1201 gcaacagcca ccgccattcg gcacagtgcc accacaaatg atgtttcctc caaactggca
     1261 gggggcagag aaggacgctg ctttcctcgc caaggacttc aactttctca ctttgaacaa
     1321 tcagccacca ccaggaaaca ggagccaacc aagggcaatg gggcccgaga acaacctgta
     1381 cagccagtac gagcagaagg tgcgcccctg cattgacctc atcgactccc tgcgggctct
     1441 gggtgtggag caggacctgg ccctgccagc catcgccgtc atcggggacc agagctcggg
     1501 caagagctct gtgctggagg cactgtcagg agtcgcgctt cccagaggca gcggaatcgt
     1561 aaccaggtgt ccgctggtgc tgaaactgaa aaagcagccc tgtgaggcat gggccggaag
     1621 gatcagctac cggaacaccg agctagagct tcaggaccct ggccaggtgg agaaagagat
     1681 acacaaagcc cagaacgtca tggccgggaa tggccggggc atcagccatg agctcatcag
     1741 cctggagatc acctcccctg aggttccaga cctgaccatc attgaccttc ccggcatcac
     1801 cagggtggct gtggacaacc agccccgaga catcggactg cagatcaagg ctctcatcaa
     1861 gaagtacatc cagaggcagc agacgatcaa cttggtggtg gttccctgta acgtggacat
     1921 tgccaccacg gaggcgctga gcatggccca tgaggtggac ccggaagggg acaggaccat
     1981 cggtatcctg accaaaccag atctaatgga caggggcact gagaaaagcg tcatgaatgt
     2041 ggtgcggaac ctcacgtacc ccctcaagaa gggctacatg attgtgaagt gccggggcca
     2101 gcaggagatc acaaacaggc tgagcttggc agaggcaacc aagaaagaaa ttacattctt
     2161 tcaaacacat ccatatttca gagttctcct ggaggagggg tcagccacgg ttccccgact
     2221 ggcagaaaga cttaccactg aactcatcat gcatatccaa aaatcgctcc cgttgttaga
     2281 aggacaaata agggagagcc accagaaggc gaccgaggag ctgcggcgtt gcggggctga
     2341 catccccagc caggaggccg acaagatgtt ctttctaatt gagaaaatca agatgtttaa
     2401 tcaggacatc gaaaagttag tagaaggaga agaagttgta agggagaatg agacccgttt
     2461 atacaacaaa atcagagagg attttaaaaa ctgggtaggc atacttgcaa ctaataccca
     2521 aaaagttaaa aatattatcc acgaagaagt tgaaaaatat gaaaagcagt atcgaggcaa
     2581 ggagcttctg ggatttgtca actacaagac atttgagatc atcgtgcatc agtacatcca
     2641 gcagctggtg gagcccgccc ttagcatgct ccagaaagcc atggaaatta tccagcaagc
     2701 tttcattaac gtggccaaaa aacattttgg cgaatttttc aaccttaacc aaactgttca
     2761 gagcacgatt gaagacataa aagtgaaaca cacagcaaag gcagaaaaca tgatccaact
     2821 tcagttcaga atggagcaga tggttttttg tcaagatcag atttacagtg ttgttctgaa
     2881 gaaagtccga gaagagattt ttaaccctct ggggacgcct tcacagaata tgaagttgaa
     2941 ctctcatttt cccagtaatg agtcttcggt ttcctccttt actgaaatag gcatccacct
     3001 gaatgcctac ttcttggaaa ccagcaaacg tctcgccaac cagatcccat ttataattca
     3061 gtattttatg ctccgagaga atggtgactc cttgcagaaa gccatgatgc agatactaca
     3121 ggaaaaaaat cgctattcct ggctgcttca agagcagagt gagaccgcta ccaagagaag
     3181 aatccttaag gagagaattt accggctcac tcaggcgcga cacgcactct gtcaattctc
     3241 cagcaaagag atccactgaa acccagcttt cttgtacaaa gtggtgatcc tctcgagcct
     3301 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     3361 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3421 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3481 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     3541 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     3601 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3661 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3721 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3781 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3841 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3901 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3961 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4021 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4081 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4141 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4201 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4261 ctgggctgtg tgcacgaanc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4321 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4381 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4441 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4501 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4561 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4621 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4681 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4741 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4801 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4861 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4921 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4981 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5041 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5101 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5161 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5221 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5281 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5341 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5401 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5461 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5521 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5581 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5641 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5701 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5761 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5821 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5881 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5941 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     6001 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6061 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6121 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6181 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     6241 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacc
//
