LOCUS       sequence-457368-        6658 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 6658)
  AUTHORS   Nishioka K, Chuikov S, Sarma K, Erdjument-Bromage H, Allis CD, 
            Tempst P, Reinberg D
  TITLE     Set9, a novel histone H3 methyltransferase that facilitates 
            transcription by precluding histone tail modifications required for 
            heterochromatin formation.
  JOURNAL   Genes Dev. 2002 Feb 15. 16(4):479-89.
  PUBMED    11850410
REFERENCE   2  (bases 1 to 6658)
  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"; journalName: "Genes Dev. 
            2002 Feb 15. 16(4):479-89."
COMMENT     Sequence Label: pcDNA Set9 wt
FEATURES             Location/Qualifiers
     source          1..6658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     215..235
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(244..672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(354..375)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     566..585
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    718..942
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     931..948
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1051..1074)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2185..2202)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2212..2229)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2248..2270
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2315..2334)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2316..2334)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2379..2582)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2408..2428)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2583..2962
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3134..3153
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3228..3332
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3333..3401,3402..4193)
                     /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     complement(3551..3570)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4364..4952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4853..4872
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5106..5123
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5240..5261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5276..5281,5282..5299,5300..5306)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5314..5330
                     /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)."
     primer_bind     5319..5341
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5338..5354
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5338..5354
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5403..5524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5414..5433)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5468..5487
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5698..6492)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5809..5828)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6419..6438
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6559..6658,1..230))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6573..6658,1..50)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6627..6646)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
       61 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
      121 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
      181 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
      241 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
      301 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
      361 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
      421 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
      481 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
      541 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
      601 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
      661 tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca
      721 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
      781 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
      841 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
      901 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcggt
      961 ttaaacttaa gcttggtacc gagctcggat ccactagtcc agtgtggtgg aattctgcag
     1021 atatccagca cagtggcggc cgcaagctta cttgtcatcg tcgtccttgt agtctccagg
     1081 acccttttgc tgggtggcct ggaaggcctt cagctccacc tggtaccact caggggcttc
     1141 aggcccactc ttcccggggg ggctgtggtc atagccatag gcaacggtga gctcttcatc
     1201 ggcctccact gctctcaggg tgcggatgca tttgatgggc ccaaaacggg ggtggacaaa
     1261 catatcgtag atgcagtttg gagtgaagga gtgatttgcc ttgtgtccca aggaggcaca
     1321 gtacttggat acgtggttat agggctcagg cacatcaatg accgtttctt catcaaggga
     1381 gagggtgttc ccattaaggg cccagtccct gctgtcaacc tcttggtgtg taattcgaac
     1441 tccattataa aaagacataa cagtattagg tcccacagct acctttgaaa aaagtccttc
     1501 tccagcactg gaaataagag attcagcaac ataaaccctt tctgattcat aaggatctgg
     1561 aagaagagca ttggtagaaa tgcaagatga agtcgactta tcaaagtggt acactgaatt
     1621 tccaggcatc agttcaaagt gaggcctccc ttcttcagtg gacataaggg tagccagttt
     1681 gccttctatc atctctccat caataaattt cccataaagt gcggtcctct catcagggta
     1741 cacataggct atcttctctc cagtcatctc cccatcttca tttacttctc ctacaaggct
     1801 tcctccatct gggtaatata tccagcacac tccatgacga atgttatctt tatactgccc
     1861 cttgaagatc agtctcccat ctgtgtcata ttcctgggct ggaccgttca gctctccgtc
     1921 tacatacgtg ccctggagaa ctcccccatc ttcgtaagtg taaactccct ggccctgcaa
     1981 ggcatcatcc acataatacc cctccagggt gctgccatca aagaagaaga acttcccccg
     2041 tccgttcttt tctccgtgaa caaagttccc ctcaaatctg tctgtggagg agtaggtgac
     2101 tgtgcagaac ccgtgcggta atccgtcatc gtccaggtgc ccttccaccg cctcctccac
     2161 catctcgtcg tcgctatcca tatggctgcc gcgcggcacc aggccgctgc tgtgatgatg
     2221 atgatgatgg ctgctgccca tggtatatct ccttcttaaa gttaaacaaa attatttcta
     2281 gagggcccgt ttaaacgcta gccagcttgg gtctccctat agtgagtcgt attaatttcg
     2341 ataagccagt aagcagtggg ttctctagtt agccagagag ctctgcttat atagacctcc
     2401 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     2461 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     2521 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     2581 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     2641 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     2701 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     2761 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     2821 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     2881 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     2941 taactagtca ataatcaatg tcaacgcgta tatctggccc gtacatcgcg aagcagcgca
     3001 aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg ccttgttgta
     3061 gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg gagcagatac
     3121 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag gggatcggga
     3181 gatctcccga tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3241 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3301 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3361 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3421 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3481 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3541 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3601 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3661 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3721 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3781 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3841 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3901 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3961 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4021 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4081 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4141 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4201 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4261 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4321 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4381 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4441 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4501 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4561 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4621 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4681 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4741 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4801 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4861 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4921 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4981 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5041 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5101 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5161 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5221 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5281 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5341 gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgacggt atacagacat
     5401 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     5461 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     5521 agttggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     5581 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     5641 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca
     5701 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     5761 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     5821 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     5881 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     5941 gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     6001 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     6061 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     6121 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     6181 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     6241 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     6301 tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     6361 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     6421 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     6481 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatccga aaatggatat
     6541 acaagctccc gggagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     6601 ctggaatagc tcagaggcag aggcggcctc ggcctctgca taaataaaaa aaattagt
//
