LOCUS       sequence-266010-        7741 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 7741)
  AUTHORS   Moberg K, Starz MA, Lees JA
  TITLE     E2F-4 switches from p130 to p107 and pRB in response to cell cycle 
            reentry.
  JOURNAL   Mol Cell Biol. 1996 Apr;16(4):1436-49.
  PUBMED    8657117
REFERENCE   2  (bases 1 to 7741)
  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: "Mol Cell 
            Biol."; date: "1996-04"; volume: "16(4)"; pages: "1436-49"
COMMENT     Sequence Label: pCMV-Neo-Bam DP1
FEATURES             Location/Qualifiers
     source          1..7741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2663..2682)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2728..2783
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2782..2801)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3141..3157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3141..3157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3154..3176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3165..3181
                     /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(3189..3195,3196..3213,3214..3219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3234..3255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3372..3389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3543..4131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3623..3642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4302..5093,5094..5162))
                     /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     4925..4944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5163..5267)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5335..5353
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5391..5413)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5513..5532
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5726..5748
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5740..5757
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5741..5757
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6168..6313
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6370..7164
                     /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(6424..6443)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7034..7053
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7358..7377)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7402..7449
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tcctcgacgg atccatggca aaagatgccg gtctaattga agccaacgga gaactcaagg
     1381 tcttcataga ccagaacctt agtcccggga aaggcgtggt gtccctcgtg gccgttcacc
     1441 cctccaccgt caacccgctc gggaagcagc tcttgccaaa aacctttgga cagtccaatg
     1501 tcaacattgc ccagcaagtg gtaattggta cgcctcagag accggcagcg tcaaacaccc
     1561 tggtggtagg aagcccacac acccccagca ctcactttgc ctctcagaac cagccttccg
     1621 actcctcacc ttggtctgcc gggaagcgca acaggaaagg agagaagaat ggcaagggcc
     1681 tacggcattt ctccatgaag gtctgcgaga aggtgcagag gaaagggacc acttcctaca
     1741 acgaagtggc agacgagctg gttgcggagt tcagtgctgc cgacaaccac atcttaccaa
     1801 acgagtcagc ttatgaccag aaaaacataa gacggcgcgt ctacgatgcc ttaaacgtgc
     1861 taatggccat gaacatcatc tccaaggaga agaaggagat caagtggatt ggtctgccca
     1921 ccaactcggc tcaggaatgt cagaacttag aggtggaaag acagaggaga cttgaaagaa
     1981 taaaacagaa acagtctcaa cttcaagaac ttattctaca gcaaattgcc ttcaagaacc
     2041 tggtgcagag aaaccggcat gcggagcagc aggccagccg gccaccgcca cccaactcag
     2101 tcatccacct gcccttcatc atcgtcaaca ccagcaagaa gacggtcatc gactgcagca
     2161 tctccaatga caaatttgag tatctgttta attttgacaa cacatttgaa atccacgatg
     2221 acatagaagt gctgaagcgg atgggcatgg cttgcgggct ggagtcgggg agctgctctg
     2281 ccgaagacct taaaatggcc agaagtctgg tccccaaggc tctggagcca tacgtgacag
     2341 aaatggctca gggaactgtt ggaggcgtgt tcatcacgac ggcaggttcc acgtctaacg
     2401 gcacaaggtt ctctgccagt gacctgacca acggtgcaga tgggatgctg gccacaagct
     2461 ccaatgggtc tcagtacagc ggctccaggg tggagactcc ggtgtcctac gtcggggagg
     2521 acgacgagga ggacgatgac ttcaacgaga atgacgagga cgactgagga tccgtcgagg
     2581 aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     2641 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     2701 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     2761 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     2821 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     2881 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     2941 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     3001 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     3061 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacggat
     3121 cggccgcaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3181 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3241 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3301 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3361 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3421 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3481 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3541 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3601 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3661 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3721 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3781 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3841 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3901 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3961 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4021 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4081 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4141 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4201 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4261 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4321 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4381 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4441 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4501 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4561 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4621 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4681 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4741 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4801 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4861 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4921 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4981 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5041 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5101 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5161 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5221 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5281 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5341 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5401 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5461 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     5521 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     5581 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5641 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     5701 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
     5761 aagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggaacacgt
     5821 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     5881 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     5941 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     6001 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     6061 tctgatggcg caggggatca agatcctgct tcatccccgt ggcccgttgc tcgcgtttgc
     6121 tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg acacaaaccc
     6181 cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg cggcgcggtc
     6241 ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca
     6301 gcgacccgct taacagcgtc aacagcgtgc cgcagatctg atcaagagac aggatgagga
     6361 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6421 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6481 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6541 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6601 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6661 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6721 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6781 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6841 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6901 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6961 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     7021 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     7081 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     7141 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     7201 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     7261 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     7321 agttcttcgc ccaccccggg agatggggga ggctaactga aacacggaag gagacaatac
     7381 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     7441 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     7501 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     7561 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aagccctgcc atagccacgg
     7621 gccccgtggg ttagggacgg cggatcgcgg ccctctagag agcttggccc attgcatacg
     7681 ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt accgccatgt
     7741 t
//
