LOCUS       Exported                8142 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pLenti6/V5-p53_R175H
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8142
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        91..121
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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     134..156
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     153..169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     153..169
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     188..208
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        190..208
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        236..462
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             463..643
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    690..815
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1308..1541
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1726..1770
                     /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        2049..2352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2353..2556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2507..2527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2553..2577
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2673..3851
                     /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             3885..3926
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3963..3983)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3968..4297
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4148..4283
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4345..4392
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4411..4809
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4896..5129
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5201..5322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5238..5257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5292..5311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5362..5497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5517..5536)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5518..5536)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5546..5563)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5546..5562)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5555..5577)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5704..6159
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5791..5810)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6001..6022
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6185..6289
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6290..7150
                     /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(6508..6527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7321..7909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7810..7829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8063..8080
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      181 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag
      241 tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta
      301 caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc
      361 ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat
      421 tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac
      481 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
      541 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
      601 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
      661 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
      721 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
      781 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
      841 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
      901 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
      961 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     1021 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     1081 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     1141 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     1201 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     1261 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     1321 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     1381 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     1441 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     1501 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     1561 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     1621 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     1681 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     1741 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     1801 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     1861 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     1921 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     1981 agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg ataagcttgg
     2041 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2101 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2161 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     2221 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     2281 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     2341 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     2401 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     2461 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     2521 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     2581 tggagacgcc atccacgctg ttttgacctc catagaagac accgactcta gaggatccac
     2641 tagtccagtg tggtggaatt gatcccttca ccatggagga gccgcagtca gatcctagcg
     2701 tcgagccccc tctgagtcag gaaacatttt cagacctatg gaaactactt cctgaaaaca
     2761 acgttctgtc ccccttgccg tcccaagcaa tggatgattt gatgctgtcc ccggacgata
     2821 ttgaacaatg gttcactgaa gacccaggtc cagatgaagc tcccagaatg ccagaggctg
     2881 ctccccgcgt ggcccctgca ccagcagctc ctacaccggc ggcccctgca ccagccccct
     2941 cctggcccct gtcatcttct gtcccttccc agaaaaccta ccagggcagc tacggtttcc
     3001 gtctgggctt cttgcattct gggacagcca agtctgtgac ttgcacgtac tcccctgccc
     3061 tcaacaagat gttttgccaa ctggccaaga cctgccctgt gcagctgtgg gttgattcca
     3121 cacccccgcc cggcacccgc gtccgcgcca tggccatcta caagcagtca cagcacatga
     3181 cggaggttgt gaggcactgc ccccaccatg agcgctgctc agatagcgat ggtctggccc
     3241 ctcctcagca tcttatccga gtggaaggaa atttgcgtgt ggagtatttg gatgacagaa
     3301 acacttttcg acatagtgtg gtggtgccct atgagccgcc tgaggttggc tctgactgta
     3361 ccaccatcca ctacaactac atgtgtaaca gttcctgcat gggcggcatg aaccggaggc
     3421 ccatcctcac catcatcaca ctggaagact ccagtggtaa tctactggga cggaacagct
     3481 ttgaggtgcg tgtttgtgcc tgtcctggga gagaccggcg cacagaggaa gagaatctcc
     3541 gcaagaaagg ggagcctcac cacgagctgc ccccagggag cactaagcga gcactgccca
     3601 acaacaccag ctcctctccc cagccaaaga agaaaccact ggatggagaa tatttcaccc
     3661 ttcagatccg tgggcgtgag cgcttcgaga tgttccgaga gctgaatgag gccttggaac
     3721 tcaaggatgc ccaggctggg aaggagccgg gggggagcag ggctcactcc agccacctga
     3781 agtccaaaaa gggtcagtct acctcccgcc ataaaaaact catgttcaag acagaagggc
     3841 ctgactcaga caagggctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta
     3901 accctctcct cggtctcgat tctacgcgta ccggttagta atgagtttgg aattaattct
     3961 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4021 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4081 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4141 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4201 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4261 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4321 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     4381 gtataatacg acaaggtgag gaactaaacc atggccaagc ctttgtctca agaagaatcc
     4441 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
     4501 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
     4561 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
     4621 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
     4681 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
     4741 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
     4801 gagggctaag cacaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt
     4861 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     4921 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     4981 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     5041 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5101 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     5161 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     5221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5281 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     5341 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5401 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5461 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     5521 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     5581 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     5641 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     5701 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5761 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5821 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5881 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5941 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6001 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6061 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6121 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     6181 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6241 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6301 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6361 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6421 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6481 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6541 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6601 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6661 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6721 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6781 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6841 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6901 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6961 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7021 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7081 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7141 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7201 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7261 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7321 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7381 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7441 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7501 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7561 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7621 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7681 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7741 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7801 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7861 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7921 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7981 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8041 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     8101 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tg
//
