LOCUS       sequence-456545-        7355 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 7355)
  AUTHORS   Trimarchi JM, Fairchild B, Wen J, Lees JA
  TITLE     The E2F6 transcription factor is a component of the mammalian 
            Bmi1-containing polycomb complex.
  JOURNAL   Proc Natl Acad Sci U S A. 2001 Feb 13;98(4):1519-24. Epub 2001 Jan 
            30.
  PUBMED    11171983
REFERENCE   2  (bases 1 to 7355)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2001-02-13"; volume: "98(4)"; pages: 
            "1519-24. Epub 2001 Jan 30"
COMMENT     Sequence Label: pCMV-Neo-Bam E2F6
FEATURES             Location/Qualifiers
     source          1..7355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     392..411
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(522..667)
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     primer_bind     complement(1078..1095)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1078..1094)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1087..1109)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1303..1322)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1422..1444
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1482..1500)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1568..1672
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1673..1741,1742..2533)
                     /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(1891..1910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2704..3292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3193..3212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3446..3463
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3580..3601
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3616..3621,3622..3639,3640..3646)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3654..3670
                     /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     3659..3681
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3678..3694
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3678..3694
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4034..4053
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4052..4107
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4153..4172
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          5181..5753
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(5844..5868)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(5865..6068)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5894..5914)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6069..6448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    6741..6788
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6813..6832
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(7026..7355,1..465))
                     /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(7137..7156)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
       61 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
      121 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
      181 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
      241 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc
      301 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
      361 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
      421 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
      481 tcctgtctct tgatcagatc tgcggcacgc tgttgacgct gttaagcggg tcgctgcagg
      541 gtcgctcggt gttcgaggcc acacgcgtca ccttaatatg cgaagtggac ctgggaccgc
      601 gccgccccga ctgcatctgc gtgttcgaat tcgccaatga caagacgctg ggcggggttt
      661 gtgtcatcat agaactaaag acatgcaaat atatttcttc cggggacacc gccagcaaac
      721 gcgagcaacg ggccacgggg atgaagcagg atcttgatcc cctgcgccat cagatccttg
      781 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
      841 cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg
      901 taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag
      961 cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtgt
     1021 tccgcttcct ttagcagccc ttgcgccctg agtgcttgcg gcagcgtgaa gcttggcact
     1081 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     1141 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     1201 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     1261 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     1321 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     1381 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     1441 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     1501 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     1561 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1621 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1681 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1741 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1801 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1861 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1921 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1981 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2041 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2101 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2161 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2221 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2281 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2341 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2401 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2461 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2521 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2581 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2641 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2701 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2761 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2821 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2881 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2941 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3001 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3061 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3121 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3181 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3241 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3301 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3361 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3421 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     3481 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3541 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     3601 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     3661 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaattgcg gccgatccgt
     3721 cgagggatct ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga
     3781 aaaatctggc tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca
     3841 caaaacaaaa tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat
     3901 ggacagcagg gggctgtttc atatactgat gacctcttta tagccaacct ttgttcatgg
     3961 cagccagcat atgggcatat gttgccaaac tctaaaccaa atactcattc tgatgtttta
     4021 aatgatttgc cctcccatat gtccttccga gtgagagaca caaaaaattc caacacacta
     4081 ttgcaatgaa aataaatttc ctttattagc cagaagtcag atgctcaagg ggcttcatga
     4141 tgtccccata atttttggca gagggaaaaa gatctcagtg gtatttgtga gccagggcat
     4201 tggccacacc agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattcctcga
     4261 cggatccatc agttgcttac ttcaagcaat tcttcacttt gctgaggatt ttcttcttcc
     4321 tcagggcctt ctggatgagt gctctcagat gaagaggtcc cgacaccttc agaccttttg
     4381 ttactggtct gaccctgctc cacttcacac aaatagacat cgataggtcc gttggtgctc
     4441 cttatgtgca ctgtgataga gtcttctctg ggagctggaa catccaatct ggtttctgct
     4501 ggagctttaa ctgcaatgac gatctgttca tggaaggcct gaatgctatg aatgtcttga
     4561 taggtcacat atgctagtct ttcattttct ttgtcatctg ttaactcaaa cagctgctga
     4621 gcacaatcct taattaactc atccaaagca tcttccattg ctgataagtc agaaagttcc
     4681 tcctgtagct tcttttgttg gggaactgct ccaaaattgc taagatcaga tcctatccat
     4741 ctaatatggt tcttggattt cttttcaacg aggtcgattc catctaagac attggtgatg
     4801 tcatacactc tccgctttcg gactcccagt ttcgttgcaa ccttgtttaa gtcaagaata
     4861 cccccgggag cagatctgac aagatccata aattttcgag ttaaataaac cagcgataca
     4921 tcaaaacgag gtctcttcac ttttagagct tttctcatgg acacatattg tacattatct
     4981 tctaaattaa tccttatttt tgatggcagc aggccctcca cgttgatggg gtctcggcac
     5041 cgacggcgaa ccgtctcctc cgtcgggtcc aggaggagac tgggaagctt cctcgccggc
     5101 cgctgctgac tcatggatcc gtcgaggaat tctttgccaa aatgatgaga cagcacaaca
     5161 accagcacgt tgcccaggag ctgtaggaaa gagaagaagg catgaacatg gttagcagag
     5221 gggcccggtt tggactcaga gtattttatc ctcatctcaa acagtgtata tcattgtaac
     5281 cataaagaga aaggcaggat gatgaccagg gtgtagttgt ttctaccaat aagaatattt
     5341 ccacgccagc cagaatttat atgcagaaat attctacctt atcatttaat tataacaatt
     5401 gttctctaaa actgtgctga agtacaatat aatataccct gattgccttg aaaaaaaagt
     5461 gattagagaa agtacttaca atctgacaaa taaacaaaag tgaatttaaa aattcgttac
     5521 aaatgcaagc taaagtttaa cgaaaaagtt acagaaaatg aaaagaaaat aagaggagac
     5581 aatggttgtc aacagagtag aaagtgaaag aaacaaaatt atcatgaggg tccatggtga
     5641 tacaagggac atcttcccat tctaaacaac accctgaaaa ctttgccccc tccatataac
     5701 atgaatttta caatagcgaa aaagaaagaa caatcaaggg tccccaaact caccctgaag
     5761 ttctcaggat cggtcgaccg gaggctggat cggtcccggt gtcttctatg gaggtcaaaa
     5821 cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct gcttatatag
     5881 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     5941 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     6001 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     6061 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     6121 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     6181 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     6241 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     6301 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     6361 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     6421 tattaataac tagtcaataa tcaatgtcaa catggcggta atgttggaca tgagccaata
     6481 taaatgtaca tattatgata tggatacaac gtatgcaatg ggccaagctc tctagagggc
     6541 cgcgatccgc cgtccctaac ccacggggcc cgtggctatg gcagggcttg ccgccccgac
     6601 gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga
     6661 aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg
     6721 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt
     6781 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     6841 cccccatctc ccggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     6901 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     6961 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     7021 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     7081 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     7141 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     7201 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     7261 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     7321 ggcgcgagcc cctgatgctc ttcgtccaga tcatc
//
