LOCUS       sequence-252070-        8142 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 8142)
  AUTHORS   Junk DJ, Vrba L, Watts GS, Oshiro MM, Martinez JD, Futscher BW
  TITLE     Different mutant/wild-type p53 combinations cause a spectrum of 
            increased invasive potential in nonmalignant immortalized human 
            mammary epithelial cells.
  JOURNAL   Neoplasia. 2008 May . 10(5):450-61.
  PUBMED    18472962
REFERENCE   2  (bases 1 to 8142)
  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: "Neoplasia. 
            2008 May . 10(5):450-61."
COMMENT     Sequence Label: pLenti6/V5-p53_R273H
FEATURES             Location/Qualifiers
     source          1..8142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        288..514
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             515..695
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    742..867
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1360..1593
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1778..1822
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     enhancer        2101..2404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2405..2608
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2559..2579
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2605..2629
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2725..3903
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVHVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     CDS             3937..3978
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(4015..4035)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4020..4349
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4200..4335
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4397..4444
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4463..4861
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4948..5181
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5253..5374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5290..5309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5344..5363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5414..5549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5569..5588)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5570..5588)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5598..5615)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5598..5614)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5607..5629)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5756..6211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5843..5862)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6053..6074
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6237..6341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6342..6410,6411..7202)
                     /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(6560..6579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7373..7961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7862..7881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8115..8132
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
      301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
      361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
      421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
      481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
      541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1981 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     2041 agacagatcc attcgattag tgaacggatc tcgacggtat cgataagctt gggagttccg
     2101 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     2161 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     2221 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     2281 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     2341 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     2401 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     2461 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     2521 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     2581 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
     2641 ccatccacgc tgttttgacc tccatagaag acaccgactc tagaggatcc actagtccag
     2701 tgtggtggaa ttgatccctt caccatggag gagccgcagt cagatcctag cgtcgagccc
     2761 cctctgagtc aggaaacatt ttcagaccta tggaaactac ttcctgaaaa caacgttctg
     2821 tcccccttgc cgtcccaagc aatggatgat ttgatgctgt ccccggacga tattgaacaa
     2881 tggttcactg aagacccagg tccagatgaa gctcccagaa tgccagaggc tgctcccccc
     2941 gtggcccctg caccagcagc tcctacaccg gcggcccctg caccagcccc ctcctggccc
     3001 ctgtcatctt ctgtcccttc ccagaaaacc taccagggca gctacggttt ccgtctgggc
     3061 ttcttgcatt ctgggacagc caagtctgtg acttgcacgt actcccctgc cctcaacaag
     3121 atgttttgcc aactggccaa gacctgccct gtgcagctgt gggttgattc cacacccccg
     3181 cccggcaccc gcgtccgcgc catggccatc tacaagcagt cacagcacat gacggaggtt
     3241 gtgaggcgct gcccccacca tgagcgctgc tcagatagcg atggtctggc ccctcctcag
     3301 catcttatcc gagtggaagg aaatttgcgt gtggagtatt tggatgacag aaacactttt
     3361 cgacatagtg tggtggtgcc ctatgagccg cctgaggttg gctctgactg taccaccatc
     3421 cactacaact acatgtgtaa cagttcctgc atgggcggca tgaaccggag gcccatcctc
     3481 accatcatca cactggaaga ctccagtggt aatctactgg gacggaacag ctttgaggtg
     3541 catgtttgtg cctgtcctgg gagagaccgg cgcacagagg aagagaatct ccgcaagaaa
     3601 ggggagcctc accacgagct gcccccaggg agcactaagc gagcactgcc caacaacacc
     3661 agctcctctc cccagccaaa gaagaaacca ctggatggag aatatttcac ccttcagatc
     3721 cgtgggcgtg agcgcttcga gatgttccga gagctgaatg aggccttgga actcaaggat
     3781 gcccaggctg ggaaggagcc aggggggagc agggctcact ccagccacct gaagtccaaa
     3841 aagggtcagt ctacctcccg ccataaaaaa ctcatgttca agacagaagg gcctgactca
     3901 gacaagggct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
     3961 ctcggtctcg attctacgcg taccggttag taatgagttt ggaattaatt ctgtggaatg
     4021 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4081 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4141 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4201 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4261 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4321 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4381 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     4441 cgacaaggtg aggaactaaa ccatggccaa gcctttgtct caagaagaat ccaccctcat
     4501 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     4561 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     4621 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     4681 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     4741 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     4801 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
     4861 agcacaattc gagctcggta cctttaagac caatgactta caaggcagct gtagatctta
     4921 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     4981 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     5041 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5101 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5161 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     5221 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     5281 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5341 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     5401 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5461 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5521 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
     5581 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     5641 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     5701 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     5761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5821 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5881 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5941 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6061 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6121 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6181 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     6241 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6301 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6361 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6421 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6481 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6541 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6601 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6661 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6721 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6781 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6841 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6901 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6961 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     7021 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7081 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7141 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     7201 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     7261 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7321 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7381 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7441 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7501 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7561 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7621 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7681 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7741 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7801 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7861 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7921 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7981 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     8041 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     8101 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//
