LOCUS       Exported                4578 bp ds-DNA     circular SYN 19-DEC-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4578)
  AUTHORS   Youjun
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Dec 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4578
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        36..1217
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     intron          266..1208
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
                     /note="color: #------"
     primer_bind     1165..1185
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
                     /note="color: black; sequence: TCAAGCCTCAGACAGTGGTTC"
     misc_feature    1252..1260
                     /label=Kozak
                     /note="color: #a6acb3"
     misc_feature    1261..1476
                     /label=hP53DD part based on addgene 25989
                     /note="color: #ffff00"
     CDS             1261..1476
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53DD
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /note="color: #993366"
                     /translation="LDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSS
                     HLKSKKGQSTSRHKKLMFKTEGPDSD"
     misc_feature    1261..1374
                     /label=Tet.
                     /note="color: #0000ff"
     primer_bind     complement(1354..1374)
                     /label=443R (Tet-rev)
                     /note="color: black; sequence: 
                     AAGGAAAAAAGCGGCCGCtcaCCCTGGCTCCTTCCCAGCCTG; added: 
                     2021-12-20"
     misc_feature    1375..1476
                     /label=3' terminal
                     /note="color: #a6acb3"
     primer_bind     complement(1599..1618)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
                     /note="color: black; sequence: TTTTGGCAGAGGGAAAAAGA"
     polyA_signal    1664..1719
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
                     /note="color: #a6acb3"
     primer_bind     complement(1718..1737)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
                     /note="color: black; sequence: CCCATATGTCCTTCCGAGTG"
     primer_bind     complement(2080..2096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(2080..2096)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     primer_bind     complement(2093..2115)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     protein_bind    2104..2120
                     /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)."
                     /note="color: #31849b"
     promoter        complement(2128..2158)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2128 .. 2134 / #ffffff / -10
                        2: 2135 .. 2152 / #ffffff
                        3: 2153 .. 2158 / #ffffff / -35"
     protein_bind    2173..2194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        2253..2448
                     /label=SV40 promoter
                     /note="SV40 early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      2299..2434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     2361..2380
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     primer_bind     complement(2384..2395)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     polyA_signal    2454..2588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(2491..2510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     2545..2564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     primer_bind     complement(2656..2673)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     rep_origin      complement(2827..3415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(2907..2926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     CDS             complement(3586..4446)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3586 .. 4377 / #ccffcc
                        2: 4378 .. 4446 / #ccffcc / signal sequence
                     Cleavage site after base 4377"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4209..4228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     promoter        complement(4447..4551)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gtcgacattg attattgact agatcatcgc gtgaggctcc ggtgcccgtc agtgggcaga
       61 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
      121 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
      181 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
      241 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
      301 ctttacgggt tatggccctt gcgtgccttg aattacttcc acgcccctgg ctgcagtacg
      361 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
      421 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
      481 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
      541 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
      601 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
      661 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
      721 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
      781 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
      841 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
      901 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
      961 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     1021 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     1081 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     1141 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     1201 catttcaggt gtcgtgagga attctgcagt cgacggtacc gcgggcccgg ggccaccatg
     1261 ctggatggag aatatttcac ccttcagatc cgtgggcgtg agcgcttcga gatgttccga
     1321 gagctgaatg aggccttgga actcaaggat gcccaggctg ggaaggagcc aggggggagc
     1381 agggctcact ccagccacct gaagtccaaa aagggtcagt ctacctcccg ccataaaaaa
     1441 ctcatgttca agacagaagg gcctgactca gactgaccgg tgtttaggcg cgccaagttg
     1501 tttaaacccg ctagcggccg cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     1561 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     1621 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     1681 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     1741 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     1801 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     1861 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     1921 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     1981 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     2041 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2101 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2161 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2221 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     2281 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2341 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2401 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     2461 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2521 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     2581 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2641 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2701 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2761 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2821 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2881 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2941 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3001 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3061 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3121 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3181 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3241 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3301 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3361 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3421 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3481 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3541 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3601 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3661 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3721 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3781 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3841 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3901 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3961 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4021 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4081 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4141 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4201 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4261 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4321 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4381 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4441 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4501 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4561 ccgaaaagtg ccacctgg
//
