LOCUS       sequence-240216-        5856 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 5856)
  AUTHORS   Zhang Y, Xiong Y, Yarbrough WG
  TITLE     ARF promotes MDM2 degradation and stabilizes p53: ARF-INK4a locus 
            deletion impairs both the Rb and p53 tumor suppression pathways.
  JOURNAL   Cell. 1998 Mar 20. 92(6):725-34.
  PUBMED    9529249
REFERENCE   2  (bases 1 to 5856)
  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: "Cell. 1998 
            Mar 20. 92(6):725-34."
COMMENT     Sequence Label: pcDNA3-myc-ARF
FEATURES             Location/Qualifiers
     source          1..5856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             897..926
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1409..1427)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1410..1427)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1447..1464)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1453..1677
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1723..2151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1810..1829)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2020..2041
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2160..2180)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2165..2494
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2345..2480
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2407..2426
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2561..3355
                     /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(2615..2634)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3225..3244
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3529..3650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3566..3585)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3620..3639
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3699..3715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3699..3715)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3712..3734)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3723..3739
                     /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(3747..3753,3754..3771,3772..3777))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3792..3813
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3930..3947)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4101..4689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4181..4200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4860..5651,5652..5720))
                     /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     5483..5502
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5721..5825)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctatggagc
      901 aaaagctcat ctcagaggaa gatctggaat tcccggggat ccccatggtg cgcaggttct
      961 tggtgaccct ccggattcgg cgcgcgtgcg gcccgccgcg agtgagggtt ttcgtggttc
     1021 acatcccgcg gctcacgggg gagtgggcag cgccaggggc gcccgccgct gtggccctcg
     1081 tgctgatgct actgaggagc cagcgtctag ggcagcagcc gcttcctaga agaccaggtc
     1141 atgatgatgg gcagcgcccg agtggcggag ctgctgctgc tccacggcgc ggagcccaac
     1201 tgcgccgacc ccgccactct cacccgaccc gtgcacgacg ctgcccggga gggcttcctg
     1261 gacacgctgg tggtgctgca ccgggccggg gcgcggctgg acgtgcgcga tgcctggggc
     1321 cgtctgcccg tggacctggc tgaggagctg ggccatcgcg atgtcgcacg gtaccaagct
     1381 tctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     1441 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1501 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1561 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     1621 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     1681 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     1741 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1801 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1861 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1921 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1981 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2041 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2101 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2161 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2221 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2281 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2341 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2401 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     2461 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     2521 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     2581 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     2641 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     2701 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     2761 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     2821 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     2881 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     2941 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3001 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3061 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3121 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3181 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3241 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3301 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3361 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3421 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3481 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     3541 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3601 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     3661 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     3721 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3781 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3841 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3901 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3961 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4021 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4081 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4141 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4201 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4261 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4321 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4381 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4441 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4501 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4561 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4621 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4681 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4741 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4801 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4861 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4921 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4981 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5041 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5101 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5161 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5221 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5281 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5341 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5401 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5461 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5521 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5581 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5641 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5701 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5761 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5821 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
