LOCUS       sequence-468752-        4745 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 4745)
  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 4745)
  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: RCAN1_pCSdest
FEATURES             Location/Qualifiers
     source          1..4745
                     /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(729..753)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    797..931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(821..840)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     875..894
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1044..1064)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1044..1062)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1083..1099)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1083..1099)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1096..1118)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1107..1123
                     /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(1131..1137,1138..1155,1156..1161))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1176..1197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1314..1331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1485..2073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1565..1584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2244..3035,3036..3104))
                     /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     2867..2886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3105..3209)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3235..3690)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3372..3393)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3584..3603
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3761..4745
                     /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 accatggagg aggtggacct gcaggacctg cccagcgcca ccatcgcctg
      181 tcacctggac ccgcgcgtgt tcgtggacgg cctgtgccgg gccaaatttg agtccctctt
      241 taggacgtat gacaaggaca tcacctttca gtattttaag agcttcaaac gagtcagaat
      301 aaacttcagc aaccccttct ccgcagcaga tgccaggctc cagctgcata agactgagtt
      361 tctgggaaag gaaatgaagt tatattttgc tcagacctta cacataggaa gctcacacct
      421 ggctccgcca aatccagaca agcagtttct gatctcccct cccgcctctc cgccagtggg
      481 atggaaacaa gtggaagatg cgaccccagt cataaactat gatctcttat atgccatctc
      541 caagctgggg ccaggggaaa agtatgaatt gcacgcagcg actgacacca ctcccagcgt
      601 ggtggtccat gtatgtgaga gtgatcaaga gaaggaggaa gaagaggaaa tggaaagaat
      661 gaggagacct aagccaaaaa ttatccagac caggaggccg gagtacacgc cgatccacct
      721 cagctagaac ccagctttct tgtacaaagt ggtgatcctc tcgagcctct agaactatag
      781 tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
      841 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      901 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      961 tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg
     1021 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     1081 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1141 gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1201 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1261 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1321 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1381 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1441 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1501 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1561 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1621 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1681 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1741 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1801 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1861 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1921 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1981 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2041 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2101 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2161 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2221 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2281 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2341 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2401 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2461 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2521 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2581 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2641 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2701 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2761 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2821 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2881 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2941 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3001 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3061 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3121 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3181 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     3241 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3301 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3361 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3421 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3481 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3541 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3601 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3661 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     3721 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc aattcaatat
     3781 ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat
     3841 ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat
     3901 ggcggacttg gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag
     3961 gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc
     4021 atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata
     4081 tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat
     4141 gtattggccc tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc
     4201 cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac
     4261 tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg
     4321 tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg
     4381 gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat
     4441 tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact
     4501 tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg
     4561 ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct
     4621 gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat
     4681 tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag
     4741 ggaac
//
