LOCUS       sequence-173999-        7715 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 7715)
  AUTHORS   Jacks T, Remington L, Williams BO, Schmitt EM, Halachmi S, Bronson 
            RT, Weinberg RA
  TITLE     Tumor spectrum analysis in p53-mutant mice.
  JOURNAL   Curr Biol. 1994 Jan 1. 4(1):1-7.
  PUBMED    7922305
REFERENCE   2  (bases 1 to 7715)
  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: "Curr Biol. 
            1994 Jan 1. 4(1):1-7."
COMMENT     Sequence Label: p53 KO neo
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(28..357)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      42..177
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(96..115)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     342..362
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(371..799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(481..502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     693..712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    845..1069
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1058..1075
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(1107..1493,1494..2108))
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKN
                     KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPT
                     EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDS
                     MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIEG
                     LLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHADA
                     LDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     2022..2041
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3499..3518)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3500..3518)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3563..3766)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3592..3612)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3767..4146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4318..4337
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4412..4516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4517..4585,4586..5377)
                     /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(4735..4754)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5548..6136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6037..6056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6290..6307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6424..6445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6460..6465,6466..6483,6484..6490)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6498..6514
                     /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     6503..6525
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6522..6538
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6522..6538
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6587..6708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6598..6617)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6652..6671
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6882..7676)
                     /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(6993..7012)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7603..7622
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 aatggatata caagctcccg ggagcttttt gcaaaagcct aggcctccaa aaaagcctcc
       61 tcactacttc tggaatagct cagaggcaga ggcggcctcg gcctctgcat aaataaaaaa
      121 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
      181 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
      241 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
      301 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
      361 ccacagaatt aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
      421 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
      481 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
      541 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
      601 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
      661 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
      721 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
      781 gcgccgctac agggcgcgtg gggatacccc ctagagcccc agctggttct ttccgcctca
      841 gaagccatag agcccaccgc atccccagca tgcctgctat tgtcttccca atcctccccc
      901 ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag acaatgcgat
      961 gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa
     1021 ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt cgaggctgat
     1081 cagcgggttt aaacgggccc tctagactac ccaccgtact cgtcaattcc aagggcatcg
     1141 gtaaacatct gctcaaactc gaagtcggcc atatccagag cgccgtaggg ggcggagtcg
     1201 tggggggtaa atcccggacc cggggaatcc ccgtccccca acatgtccag atcgaaatcg
     1261 tctagcgcgt cggcatgcgc catcgccacg tcctcgccgt ctaagtggag ctcgtccccc
     1321 aggctgacat cggtcggggg ggccgtcgac agtctgcgcg tgtgtcccgc ggggagaaag
     1381 gacaggcgcg gagccgccag ccccgcctct tcgggggcgt cgtcgtccgg gagatcgagc
     1441 aggccctcga tggtagaccc gtaattgttt ttcgtacgcg cgcggctgta cgcggaccca
     1501 ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc gaataagaag
     1561 gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata
     1621 ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat
     1681 acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa
     1741 aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta
     1801 ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa
     1861 cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga
     1921 gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt
     1981 acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt
     2041 aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta
     2101 tctaatctta gctggaagta caggttctca ccggtcgatg aagtgtcctt ggccagggag
     2161 gagctggcgg tggtgcagga ctcatcttgg ggacccaggc tgggtggggt gcgtccaccg
     2221 gtatcgatga gggcgtcatg caggtcatgg tccattgtac tctgtgtata agagcttgag
     2281 ggtgtgtccg cgctgtagat ctggagaagc ccgcaatatt gacacaacac aaagagaccc
     2341 cggcaggtct taaagagctg caaaatgtac tttctgaact tttcgccaag gaaaaagtag
     2401 ataattggat taagacaaca gtgtacgaag gcaagtgtct cggtagcctg gatggcgtaa
     2461 tccaggtagc gctcgaaggt gcaatcctgc agtacttcga gttctaccag tgtttccaaa
     2521 aagagtacta tgttataagg tgtccagaac cccaggaaca gtactaccac tgcgaagatc
     2581 atcttgaccg ccttgttttt tttctcgttt ttacaatgct gcagtgttct aattatcatg
     2641 ctgtagcaaa agagcatgat gcccagcgga atcacaagcc ccaggatatt aatctccaga
     2701 ctagaaagca ccttccatgt tgtgctattg aggctgtact tggtcttaca atacgtatgg
     2761 ttgcgctctg tgtagcacgt gctgaagagg aatccgggga gtgaggcgaa aactgccact
     2821 gaccacgtgg ccagggaggt aataacccca taggtgagag tccttgcccg aagagaaaag
     2881 acagcatgca caatggccaa atatctatca atagacatga gcatcacgaa aaagatccct
     2941 gagtagaacc ccacaaggta catccaagaa atcatcttac acaggccgag tccgaacacc
     3001 cactggtcgg ccgcataata cccccaaaag ggcagggaaa atacgaaaag aagatcgcta
     3061 attgccagat tcagcaagta aacgtcggtc atgctcctaa gtctcttgta cttgaacaat
     3121 accaatacca caacggagtt acccaacaag ccgaagacaa acaccaggga gtacaagggg
     3181 gggaggaaaa gctccccgaa cgctttaatg ccctccttag tgcacggctt tgggatgctt
     3241 tcgtacaagt agtaattgct gtagatactc tcgtccaaag tggtatctgc gatgtcagtg
     3301 ggattcatat cgatgctggc gtcatcatcg tccttgtagt cggcgaatac caggcagaag
     3361 atgtagctca gggcgatgat cgtcttcatg gtggcgcggc cgccactgtg ctggatatct
     3421 gcagaattcc accacactgg actagtggat ccgagctcgg taccaagctt aagtttaaac
     3481 gctagccagc ttgggtctcc ctatagtgag tcgtattaat ttcgataagc cagtaagcag
     3541 tgggttctct agttagccag agagctctgc ttatatagac ctcccaccgt acacgcctac
     3601 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     3661 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     3721 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     3781 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     3841 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     3901 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     3961 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     4021 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     4081 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc
     4141 aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc ctaaccctaa
     4201 gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa attttgctcg
     4261 cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt aactatgcgg
     4321 catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc ccgatccgtc
     4381 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4441 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4501 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4561 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4621 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4681 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4741 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4801 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4861 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4921 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4981 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5041 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5101 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5161 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5221 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5281 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5341 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5401 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5461 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6421 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6481 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     6541 tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag atacattgat
     6601 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     6661 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg ggtgggcgaa
     6721 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     6781 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     6841 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     6901 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     6961 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     7021 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     7081 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     7141 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     7201 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     7261 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     7321 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     7381 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     7441 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     7501 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     7561 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     7621 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     7681 aacgatcctc atcctgtctc ttgatcagat ccgaa
//
