LOCUS       sequence-169367-        7510 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 7510)
  TITLE     Sidransky Plasmids
REFERENCE   2  (bases 1 to 7510)
  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"
COMMENT     Sequence Label: TAp63alpha-FLAG
FEATURES             Location/Qualifiers
     source          1..7510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2858..2881
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2932..2949)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2938..3162
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3208..3636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3295..3314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3505..3526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3645..3665)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3650..3979
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3830..3965
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3892..3911
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4028..5053
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4028..5053
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5183..5304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5220..5239)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5274..5293
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5353..5369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5353..5369)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5366..5388)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5377..5393
                     /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)."
     promoter        complement(join(5401..5407,5408..5425,5426..5431))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5446..5467
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5584..5601)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5755..6343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5835..5854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6514..7305,7306..7374))
                     /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     7137..7156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7375..7479)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccatgtcc cagagcacac agacaaatga
      961 attcctcagt ccagaggttt tccagcatat ctgggatttt ctggaacagc ctatatgttc
     1021 agttcagccc attgacttga actttgtgga tgaaccatca gaagatggtg cgacaaacaa
     1081 gattgagatt agcatggact gtatccgcat gcaggactcg gacctgagtg accccatgtg
     1141 gccacagtac acgaacctgg ggctcctgaa cagcatggac cagcagattc agaacggctc
     1201 ctcgtccacc agtccctata acacagacca cgcgcagaac agcgtcacgg cgccctcgcc
     1261 ctacgcacag cccagctcca ccttcgatgc tctctctcca tcacccgcca tcccctccaa
     1321 caccgactac ccaggcccgc acagtttcga cgtgtccttc cagcagtcga gcaccgccaa
     1381 gtcggccacc tggacgtatt ccactgaact gaagaaactc tactgccaaa ttgcaaagac
     1441 atgccccatc cagatcaagg tgatgacccc acctcctcag ggagctgtta tccgcgccat
     1501 gcctgtctac aaaaaagctg agcacgtcac ggaggtggtg aagcggtgcc ccaaccatga
     1561 gctgagccgt gaattcaacg agggacagat tgcccctcct agtcatttga ttcgagtaga
     1621 ggggaacagc catgcccagt atgtagaaga tcccatcaca ggaagacaga gtgtgctggt
     1681 accttatgag ccaccccagg ttggcactga attcacgaca gtcttgtaca atttcatgtg
     1741 taacagcagt tgtgttggag ggatgaaccg ccgtccaatt ttaatcattg ttactctgga
     1801 aaccagagat gggcaagtcc tgggccgacg ctgctttgag gcccggatct gtgcttgccc
     1861 aggaagagac aggaaggcgg atgaagatag catcagaaag cagcaagttt cggacagtac
     1921 aaagaacggt gatggcacga agcgcccgtt tcgtcagaac acacatggta tccagatgac
     1981 atccatcaag aaacgaagat ccccagatga tgaactgtta tacttaccag tgaggggccg
     2041 tgagacttat gaaatgctgt tgaagatcaa agagtccctg gaactcatgc agtaccttcc
     2101 tcagcacaca attgaaacgt acaggcaaca gcaacagcag cagcaccagc acttacttca
     2161 gaaacagacc tcaatacagt ctccatcttc atatggtaac agctccccac ctctgaacaa
     2221 aatgaacagc atgaacaagc tgccttctgt gagccagctt atcaaccctc agcagcgcaa
     2281 cgccctcact cctacaacca ttcctgatgg catgggagcc aacattccca tgatgggcac
     2341 ccacatgcca atggctggag acatgaatgg actcagcccc acccaggcac tccctccccc
     2401 actctccatg ccatccacct cccactgcac acccccacct ccgtatccca cagattgcag
     2461 cattgtcagt ttcttagcga ggttgggctg ttcatcatgt ctggactatt tcacgaccca
     2521 ggggctgacc accatctatc agattgagca ttactccatg gatgatctgg caagtctgaa
     2581 aatccctgag caatttcgac atgcgatctg gaagggcatc ctggaccacc ggcagctcca
     2641 cgaattctcc tccccttctc atctcctgcg gaccccaagc agtgcctcta cagtcagtgt
     2701 gggctccagt gagacccggg gtgagcgtgt tattgatgct gtgcgattca ccctccgcca
     2761 gaccatctct ttcccacccc gagatgagtg gaatgacttc aactttgaca tggatgctcg
     2821 ccgcaataag caacagcgca tcaaagagga gggggaggac tacaaggacg acgatgacaa
     2881 gtagtaggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     2941 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3001 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3061 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3121 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3181 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3241 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3301 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3361 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3421 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3481 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3541 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3601 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3661 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3721 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3781 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3841 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3901 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3961 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4021 gcacgtgatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     4081 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     4141 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4201 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     4261 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     4321 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     4381 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4441 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     4501 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     4561 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     4621 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     4681 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     4741 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     4801 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     4861 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     4921 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     4981 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     5041 ggcaaaggaa tagcacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
     5101 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5161 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5221 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5281 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5341 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5401 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5461 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5521 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5581 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5641 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5701 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5761 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5821 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5881 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5941 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6001 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6061 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6181 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6301 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6361 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6421 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6481 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6541 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6601 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6661 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6721 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6781 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6841 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6901 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6961 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7021 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7081 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7141 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7201 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7261 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7321 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7381 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7441 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7501 acctgacgtc
//
