LOCUS       sequence-473533-        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_R175H
FEATURES             Location/Qualifiers
     source          1..8142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             180..413
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    485..606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(522..541)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     576..595
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      646..781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(801..820)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(802..820)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(830..847)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(830..846)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(839..861)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      988..1443
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1075..1094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1285..1306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1469..1573
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1574..1642,1643..2434)
                     /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(1792..1811)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2605..3193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3094..3113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3347..3364
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3481..3502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3517..3522,3523..3540,3541..3547)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3555..3571
                     /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     3560..3582
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3579..3595
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3579..3595
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3614..3634
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3616..3634
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3662..3888
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3889..4069
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4116..4241
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4734..4967
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5152..5196
                     /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        5475..5778
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5779..5982
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5933..5953
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5979..6003
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             6099..7277
                     /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
                     SPDDIEQWFTEDPGPDEAPRMPEAAPRVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRHCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     CDS             7311..7352
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(7389..7409)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7394..7723
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7574..7709
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        7771..7818
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(7837..8142,1..93)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa
       61 ttgctgccct ctggttatgt gtgggagggc taagcacaat tcgagctcgg tacctttaag
      121 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
      181 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
      421 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
      481 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
      541 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
      601 atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact
      661 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
      721 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
      781 ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
      841 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
      901 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      961 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     1021 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1081 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1141 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1201 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1261 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1321 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1381 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     1441 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1501 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1561 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1621 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1681 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1741 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1801 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1861 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1921 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1981 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2041 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2101 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2161 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2221 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2281 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2341 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2401 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2461 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2521 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2581 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2641 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2701 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2761 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2821 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2881 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2941 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3001 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3061 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3121 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3181 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3241 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3301 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3361 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3421 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3481 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     3541 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     3601 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta
     3661 atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
     3721 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
     3781 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
     3841 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
     3901 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     3961 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     4021 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     4081 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     4141 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     4201 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     4261 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     4321 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     4381 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     4441 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     4501 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     4561 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     4621 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     4681 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     4741 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     4801 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     4861 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     4921 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     4981 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     5041 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     5101 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     5161 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     5221 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     5281 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     5341 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     5401 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgataa
     5461 gcttgggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     5521 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     5581 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     5641 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     5701 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     5761 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     5821 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     5881 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     5941 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
     6001 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg actctagagg
     6061 atccactagt ccagtgtggt ggaattgatc ccttcaccat ggaggagccg cagtcagatc
     6121 ctagcgtcga gccccctctg agtcaggaaa cattttcaga cctatggaaa ctacttcctg
     6181 aaaacaacgt tctgtccccc ttgccgtccc aagcaatgga tgatttgatg ctgtccccgg
     6241 acgatattga acaatggttc actgaagacc caggtccaga tgaagctccc agaatgccag
     6301 aggctgctcc ccgcgtggcc cctgcaccag cagctcctac accggcggcc cctgcaccag
     6361 ccccctcctg gcccctgtca tcttctgtcc cttcccagaa aacctaccag ggcagctacg
     6421 gtttccgtct gggcttcttg cattctggga cagccaagtc tgtgacttgc acgtactccc
     6481 ctgccctcaa caagatgttt tgccaactgg ccaagacctg ccctgtgcag ctgtgggttg
     6541 attccacacc cccgcccggc acccgcgtcc gcgccatggc catctacaag cagtcacagc
     6601 acatgacgga ggttgtgagg cactgccccc accatgagcg ctgctcagat agcgatggtc
     6661 tggcccctcc tcagcatctt atccgagtgg aaggaaattt gcgtgtggag tatttggatg
     6721 acagaaacac ttttcgacat agtgtggtgg tgccctatga gccgcctgag gttggctctg
     6781 actgtaccac catccactac aactacatgt gtaacagttc ctgcatgggc ggcatgaacc
     6841 ggaggcccat cctcaccatc atcacactgg aagactccag tggtaatcta ctgggacgga
     6901 acagctttga ggtgcgtgtt tgtgcctgtc ctgggagaga ccggcgcaca gaggaagaga
     6961 atctccgcaa gaaaggggag cctcaccacg agctgccccc agggagcact aagcgagcac
     7021 tgcccaacaa caccagctcc tctccccagc caaagaagaa accactggat ggagaatatt
     7081 tcacccttca gatccgtggg cgtgagcgct tcgagatgtt ccgagagctg aatgaggcct
     7141 tggaactcaa ggatgcccag gctgggaagg agccgggggg gagcagggct cactccagcc
     7201 acctgaagtc caaaaagggt cagtctacct cccgccataa aaaactcatg ttcaagacag
     7261 aagggcctga ctcagacaag ggctcgagtc tagagggccc gcggttcgaa ggtaagccta
     7321 tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg ttagtaatga gtttggaatt
     7381 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     7441 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     7501 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     7561 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     7621 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     7681 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     7741 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     7801 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa
     7861 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     7921 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     7981 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     8041 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     8101 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ct
//
