LOCUS       Exported                4676 bp ds-DNA     circular SYN 19-DEC-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4676)
  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..4676
                     /organism="synthetic DNA construct"
                     /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    1263..1271
                     /label=Kozak sequence
                     /note="color: #a6acb3"
     CDS             1272..1574
                     /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="EPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERF
                     EMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     misc_feature    1272..1358
                     /label=NLS
                     /note="color: #a6acb3"
     misc_feature    1290..1574
                     /label=hP53DD part based on addgene 25989
                     /note="color: #ffff00"
     misc_feature    1359..1472
                     /label=Tet.
                     /note="color: #0000ff"
     misc_feature    1473..1574
                     /label=3' terminal
                     /note="color: #a6acb3"
     primer_bind     complement(1697..1716)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
                     /note="color: black; sequence: TTTTGGCAGAGGGAAAAAGA"
     polyA_signal    1762..1817
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
                     /note="color: #a6acb3"
     primer_bind     complement(1816..1835)
                     /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(2178..2194)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(2178..2194)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     primer_bind     complement(2191..2213)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     protein_bind    2202..2218
                     /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(2226..2256)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2226 .. 2232 / #ffffff / -10
                        2: 2233 .. 2250 / #ffffff
                        3: 2251 .. 2256 / #ffffff / -35"
     protein_bind    2271..2292
                     /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        2351..2546
                     /label=SV40 promoter
                     /note="SV40 early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      2397..2532
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     2459..2478
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     primer_bind     complement(2482..2493)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     polyA_signal    2552..2686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(2589..2608)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     2643..2662
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     primer_bind     complement(2754..2771)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     rep_origin      complement(2925..3513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(3005..3024)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     CDS             complement(3684..4544)
                     /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: 3684 .. 4475 / #ccffcc
                        2: 4476 .. 4544 / #ccffcc / signal sequence
                     Cleavage site after base 4475"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4307..4326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     promoter        complement(4545..4649)
                     /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 gatccaccgg
     1261 tcgccaccat ggagcctcac cacgagctgc ccccagggag cactaagcga gcactgccca
     1321 acaacaccag ctcctctccc cagccaaaga agaaaccact ggatggagaa tatttcaccc
     1381 ttcagatccg tgggcgtgag cgcttcgaga tgttccgaga gctgaatgag gccttggaac
     1441 tcaaggatgc ccaggctggg aaggagccag gggggagcag ggctcactcc agccacctga
     1501 agtccaaaaa gggtcagtct acctcccgcc ataaaaaact catgttcaag acagaagggc
     1561 ctgactcaga ctgaccggtg tttaggcgcg ccaagttgtt taaacccgct agcggccgca
     1621 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     1681 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     1741 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     1801 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     1861 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     1921 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     1981 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     2041 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     2101 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     2161 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2221 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2281 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2341 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2401 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2461 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     2521 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     2581 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2641 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     2701 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2761 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2821 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2881 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2941 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3001 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3061 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3121 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3181 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3241 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3301 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3361 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3421 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3481 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3541 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3601 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3661 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3721 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3781 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3841 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3901 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3961 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4021 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4081 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4141 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4201 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4261 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4321 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4381 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4441 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4501 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4561 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4621 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgg
//
