LOCUS       sequence-324480-       10109 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 10109)
  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 10109)
  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: LTN1_pCSdest
FEATURES             Location/Qualifiers
     source          1..10109
                     /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(7071..7095)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    7139..7273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7163..7182)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7217..7236
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7386..7406)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7386..7404)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7425..7441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7425..7441)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7438..7460)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7449..7465
                     /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(7473..7479,7480..7497,7498..7503))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7518..7539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7656..7673)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7827..8415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7907..7926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8586..9377,9378..9446))
                     /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     9209..9228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9447..9551)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9577..10032)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9714..9735)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9926..9945
                     /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 aagcaggctt taaaggaacc aattcagtcg acccacgcgt
     1141 ccgaaccggg gtctgagtgc tcagagtaca gctgcaaccg cgaccatggg cgggaagaac
     1201 aagcagcgaa ctaaagggaa cctgaggcct tcaaacagtg gccgagctgc agaactcctt
     1261 gccaaagaac agggaacagt gcctggattt attggttttg gaacatctca gagtgaccta
     1321 ggctatgttc ctgctattca aggagctgaa gaaattgaca gtcttgtaga ttctgatttc
     1381 cgaatggtgc tgcggaaact ttcaaagaaa gatgtcacca caaaattaaa agctatgcag
     1441 gaatttggaa ccatgtgtac agagagagac acagaaactg tgaaaggagt tcttccatat
     1501 tggccaagaa ttttttgcaa aatttcactt gatcatgacc gtcgcgtccg agaagccaca
     1561 caacaagctt ttgaaaaact tatccttaaa gtaaagaaac agttggctcc ctacttaaaa
     1621 agtttaatgg gatattggct aatggctcag tgtgatactt acacaccagc tgcgtttgca
     1681 gcaaaagatg catttgaagc ggcttttcct ccaagcaagc aacctgaagc catagcattt
     1741 tgtaaggatg aaattacaag tgtgctgcag gatcatctta taaaagaaac acctgataca
     1801 ctcagtgacc cgcaaactgt tccagaggaa gaaagagaag ctaaattcta ccgggttgta
     1861 acttgttcct tattggcatt aaagagatta ctttgccttt tacctgataa tgagcttgat
     1921 tctctggagg agaaatttaa gtctctttta tcacagaata agttttggaa gtatggaaaa
     1981 cacagtgtac ctcagattcg ctcagcttat tttgagttag tctctgcatt gtgccagcgc
     2041 attccacagt tgatgaaaga ggaagcatcc aaagtgagcc catcagttct acttagcatt
     2101 gatgacagtg acccaattgt ctgcccagct ctctgggaag ctgtactcta tacacttaca
     2161 actattgagg actgttggct tcatgtaaat gcaaaaaaga gtgtgtttcc caagctatca
     2221 actgtgattc gtgaaggtgg tcggggtcta gctactgtca tatatcctta ccttctgcca
     2281 ttcatcagca agctccctca gtccatcaca aatccaaagt tggatttctt caaaaatttc
     2341 ctcacgtctc tagttgctgg gctgtcaaca gagagaacta aaaccagctc ttcagagtcc
     2401 tcggcagtaa tatctgcttt ttttgaatgc ttacgtttta taatgcagca aaacttaggt
     2461 gaggaagaga ttgaacagat gctcgtcaat gatcagttga tcccttttat tgatgcagtt
     2521 ctcaaagacc caggattgca acatgggcag ctatttaacc atttagcaga aactctaagt
     2581 tcctgggaag ccaaagcaga cacggaaaaa gatgaaaaaa cagctcacaa cttggagaac
     2641 gtactgatac atttctggga aagactgtca gagatctgtg ttgcgaaaat cagtgagcca
     2701 gaagctgatg ttgagtccgt tttgggtgta tctaacctat tacaggtgct tcagaagccg
     2761 aagagctcat tgaagtcaag taaaaaaaaa aatggtaagg ttagatttgc tgatgagata
     2821 cttgaaagca ataaagagaa tgaaaaatgt gtatcttcag aaggagagaa gattgaaggc
     2881 tgggaattaa caactgaacc ttctctcact cataattctt caggcctttt gtctcctcta
     2941 aggaaaaaac ctttggaaga cttagtctgt aaactcgcag atataagtat taattatgtc
     3001 aatgaacgaa agtcagagca acatctaagg tttctttcta ctctgcttga ctccttttct
     3061 tcaagccgag tatttaaaat gctacttggt gatgaaaaac agagtattgt ccaagccaaa
     3121 cctcttgaaa tagccaagct tgtacaaaaa aatcctgcgg tgcagttttt ataccagaaa
     3181 ctgataggtt ggctaaatga agatcaaagg aaggattttg gtttcctggt ggacattttg
     3241 tacagtgctc tccggtgctg tgacaatgat atggaaagaa aaaaagtctt ggatgatcta
     3301 accaaggtgg acttgaaatg gaattctctt cttaagatta ttgaaaaggc atgtcctagt
     3361 tcagataaac atgctttagt aactccttgg ctcaaaggcg atatccttgg tgagaaattg
     3421 gtcaacttgg cagattgtct ttgtaatgag gacttggaat ccagggtatc ttcagaatct
     3481 cacttctcag aaagatggac tcttctaagc ttggtattat cccaacatgt taaaaatgat
     3541 tacttgattg gagacgtata tgttgaaaga atcattgtta gacttcatga aactttattc
     3601 aaaacaaaga aattatcaga agctgaaagc agtgactcat cagtgtcttt tatctgtgat
     3661 gtggcctata actatttcag ctcagcgaaa ggatgcttgc taatgccatc atctgaagat
     3721 ttattattaa ctctctttca gttatgtgct cagagcaaag aaaaaacaca tttgccagat
     3781 tttcttatct gtaaactgaa aaatacttgg ctctctggtg taaatttatt ggttcatcaa
     3841 actgacagtt catataaaga gagtaccttc ctacatttgt ctgctctgtg gctgaagaac
     3901 caagttcagg cttcatcttt ggatatcaac agtctccaag tcctcttgtc tgctgttgat
     3961 gatttgctaa atacacttct agagagtgaa gattcttatc ttatgggagt ttatattgga
     4021 agtgtaatgc cgaacgacag tgaatgggaa aagatgaggc agtctcttcc tatgcagtgg
     4081 ttacatagac ctcttttaga gggaagattg agtttgaatt atgaatgttt caaaacagat
     4141 tttaaggaac aggacataaa gacacttccc agccatttgt gtacttcagc attattgagc
     4201 aaaatggtct taattgcact gagaaaggaa acagtcttag aaaataatga gcttgagaaa
     4261 ataattgcag aactgcttta ttcactgcag tggtgtgaag aattagataa cccacctatt
     4321 tttctaattg gattttgtga aatacttcaa aaaatgaata ttacgtatga taacttacgt
     4381 gtacttggta atacgtcggg ccttttgcag ctgttattta acaggtccag agaacatggc
     4441 acactgtggt ctcttattat tgctaagttg atcctttccc gaagcatttc atctgatgaa
     4501 gtaaaaccac attataagag aaaagaaagt ttttttccac taactgaagg caatttgcat
     4561 accattcaaa gtctatgtcc atttttgtca aaagaagaaa agaaagaatt tagtgctcaa
     4621 tgtatacctg ctcttttggg ctggactaag aaagatcttt gcagcactaa tggaggtttt
     4681 ggacatcttg ccattttcaa ttcttgtctg caaaccaaaa gtatagatga tggagagcta
     4741 ttacatggaa tattaaaaat cataatatcc tggaagaaag agcatgaaga tatttttctt
     4801 ttcagttgta atctatcaga agcaagtcca gaggtactgg gtgtaaatat agaaataatc
     4861 cggtttcttt ccctatttct gaaatactgc tcatcccctt tggcagagag tgagtgggac
     4921 ttcatcatgt gctccatgtt ggcttggttg gagacaacaa gtgagaatca ggcattgtat
     4981 tctattccac ttgtgcaact gtttgcctgt gtcagctgtg atttggcctg tgacctcagt
     5041 gctttctttg attccacaac tctggatacc attggcaatc ttcctgtaaa tctaatcagt
     5101 gaatggaaag aatttttttc ccaaggcatc cacagtttgc ttttacctat tttggtgact
     5161 gttacaggag aaaacaaaga tgtgtctgaa acatcctttc agaatgcaat gctgaaaccc
     5221 atgtgtgaaa cattaacgta tatctcaaag gaacagctat tgagtcacaa acttcctgca
     5281 agattagttg ctgaccaaaa aacaaactta ccagaatatc tccagacttt gttaaataca
     5341 ttggccccat tactcctctt cagagctagg cctgtgcaaa ttgctgttta tcatatgcta
     5401 tacaaattga tgcctgaatt accacagtat gatcaggata atctaaagtc atacggagat
     5461 gaagaagaag agccagcctt gtcaccacca gcagcactga tgtctcttct tagcattcaa
     5521 gaggacttac tagaaaatgt tttggggtgt attcctgttg gacagatagt tactattaaa
     5581 ccactgagtg aagacttctg ttatgttctg ggataccttc tcacttggaa attaatacta
     5641 actttcttca aagctgcatc atcacagctt cgggctttgt attccatgta tcttcggaaa
     5701 acaaagagtt tgaataaatt gctctatcac ctgttcaggc ttatgccaga aaatccaacc
     5761 tatgcagaga cagcagttga ggtcccaaat aaggacccta aaacattctt tactgaggag
     5821 ctccagctga gtattagaga aacaacaatg cttccatacc acattccaca cttggcttgt
     5881 tcagtctatc atatgacatt aaaagacttg cctgccatgg ttaggttgtg gtggaatagc
     5941 agtgagaagc gtgttttcaa tattgtggat agatttacaa gcaagtatgt cagcagtgtt
     6001 ctttcttttc aagaaatatc ttctgtacaa acaagtacac aactatttaa tggcatgacg
     6061 gttaaagctc gagctactac tcgagaggta atggctactt atactattga ggacatagtt
     6121 attgaactta taatacaact gccttcaaat tatccactgg gttcaataat agtagaaagt
     6181 gggaaaagag taggagtagc tgttcagcag tggcggaact ggatgctgca gttaagcact
     6241 tacctcaccc atcagaatgg aagtattatg gaaggcttag ctttatggaa aaataacgta
     6301 gacaaacgtt ttgagggtgt tgaagattgc atgatctgtt tctcagtcat tcacggtttc
     6361 aactattccc ttcccaaaaa agcctgtaga acatgcaaga aaaaattcca ttcagcctgc
     6421 ttgtacaaat ggtttacatc tagcaacaaa tccacttgtc cactgtgtcg tgagacgttt
     6481 ttctgagatt tttttcactg gaagggatcc ctgaagtaca tcaaacaaag gcattggatt
     6541 tggatccgtc tgaaagtgtg gatgtgggga agccagtgag cattactttt aaataggacc
     6601 ttctctggaa aattattttg gttaatgtaa taatcctaaa atcaggttta cttaacttta
     6661 gattgctttg taaatgtttg gaaactttct tttacaaaag aatattacat atttgagaga
     6721 aaatatttct attaatggtt taatttcttt gtatatttaa aaataaatat gaaaaacccc
     6781 taaattacag aattggaatt tgtgaaaaga ttgcacaact atattattga taaacccatt
     6841 ccgtttgttg atctgtgttt tagaatagtc caagtgaagc aaaacgaact gagagagctg
     6901 atttacatct atagatattt taagccttat ttattagtta tgacaataaa ttagtattct
     6961 gatatttgta gattatttta atgataaaaa tgacactaag ctattaatac ggcttgtctt
     7021 aagatatctg gaggcatgct ctgaaatcct atataaaaga gtttatctag acccagcttt
     7081 cttgtacaaa gtggtgatcc tctcgagcct ctagaactat agtgagtcgt attacgtaga
     7141 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     7201 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     7261 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     7321 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     7381 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     7441 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt
     7501 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7561 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7621 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7681 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7741 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7801 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7861 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7921 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7981 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     8041 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     8101 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8161 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8221 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8281 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8341 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8401 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8461 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8521 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8581 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8641 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8701 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8761 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8821 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8881 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8941 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     9001 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     9061 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     9121 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9181 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     9241 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9301 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9361 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9421 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9481 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9541 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     9601 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     9661 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     9721 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     9781 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     9841 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     9901 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     9961 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
    10021 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
    10081 gggcctcttc gctattacgc cagtcgacc
//
