LOCUS       sequence-63873-e        6841 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 6841)
  AUTHORS   Yeh E, Cunningham M, Arnold H, Chasse D, Monteith T, Ivaldi G, Hahn 
            WC, Stukenberg PT, Shenolikar S, Uchida T, Counter CM, Nevins JR, 
            Means AR, Sears R
  TITLE     A signalling pathway controlling c-Myc degradation that impacts 
            oncogenic transformation of human cells.
  JOURNAL   Nat Cell Biol. 2004 Apr;6(4):308-18. Epub 2004 Mar 14.
  PUBMED    15048125
REFERENCE   2  (bases 1 to 6841)
  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: "Nat Cell 
            Biol."; date: "2004-04"; volume: "6(4)"; pages: "308-18. Epub 2004 
            Mar 14"
COMMENT     Sequence Label: pD40-His/V5-c-Myc
FEATURES             Location/Qualifiers
     source          1..6841
                     /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"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2289..2313)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2339..2380
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2390..2407
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2430..2447)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2436..2660
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2706..3134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2793..2812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3003..3024
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3143..3163)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3148..3477
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3328..3463
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3390..3409
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3544..4338
                     /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(3598..3617)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4208..4227
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4514..4635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4551..4570)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4605..4624
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4684..4700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4684..4700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4697..4719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4708..4724
                     /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(4732..4738,4739..4756,4757..4762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4777..4798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4915..4932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5086..5674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5166..5185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5845..6636,6637..6705))
                     /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     6468..6487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6706..6810)
                     /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 gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggctccgcg gccgccccct tcaccatgcc
      961 cctcaacgtg aacttcacca acaggaacta tgacctcgac tacgactccg tacagcccta
     1021 tttcatctgc gacgaggaag agaatttcta tcaccagcaa cagcagagcg agctgcagcc
     1081 gcccgcgccc agtgaggata tctggaagaa attcgagctg cttcccaccc cgcccctgtc
     1141 cccgagccgc cgctccgggc tctgctctcc atcctatgtt gcggtcgcta cgtccttctc
     1201 cccaagggaa gacgatgacg gcggcggtgg caacttctcc accgccgatc agctggagat
     1261 gatgaccgag ttacttggag gagacatggt gaaccagagc ttcatctgcg atcctgacga
     1321 cgagaccttc atcaagaaca tcatcatcca ggactgtatg tggagcggtt tctcagccgc
     1381 tgccaagctg gtctcggaga agctggcctc ctaccaggct gcgcgcaaag acagcaccag
     1441 cctgagcccc gcccgcgggc acagcgtctg ctccacctcc agcctgtacc tgcaggacct
     1501 caccgccgcc gcgtccgagt gcattgaccc ctcagtggtc tttccctacc cgctcaacga
     1561 cagcagctcg cccaaatcct gtacctcgtc cgattccacg gccttctctc cttcctcgga
     1621 ctcgctgctg tcctccgagt cctccccacg ggccagccct gagcccctag tgctgcatga
     1681 ggagacaccg cccaccacca gcagcgactc tgaagaagag caagaagatg aggaagaaat
     1741 tgatgtggtg tctgtggaga agaggcaaac ccctgccaag aggtcggagt cgggctcatc
     1801 tccatcccga ggccacagca aacctccgca cagcccactg gtcctcaaga ggtgccacgt
     1861 ctccactcac cagcacaact acgccgcacc cccctccaca aggaaggact atccagctgc
     1921 caagagggcc aagttggaca gtggcagggt cctgaagcag atcagcaaca accgcaagtg
     1981 ctccagcccc aggtcctcag acacggagga aaacgacaag aggcggacac acaacgtctt
     2041 ggaacgtcag aggaggaacg agctgaagcg cagctttttt gccctgcgtg accagatccc
     2101 tgaattggaa aacaacgaaa aggcccccaa ggtagtgatc ctcaaaaaag ccaccgccta
     2161 catcctgtcc attcaagcag acgagcacaa gctcacctct gaaaaggact tattgaggaa
     2221 acgacgagaa cagttgaaac acaaactcga acagcttcga aactctggtg caaagggtgg
     2281 gcgcgccgac ccagctttct tgtacaaagt ggttgatcta gagggcccgc ggttcgaagg
     2341 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca
     2401 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2461 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2521 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2581 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2641 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2701 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2761 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2821 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2881 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2941 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3001 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3061 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3121 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3241 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3301 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3361 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3421 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3481 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3541 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3601 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3661 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3721 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3781 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3841 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3901 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3961 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4021 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4081 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4141 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4201 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4261 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4321 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     4381 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4441 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4501 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4561 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4621 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6841 c
//
