LOCUS       sequence-313557-        7345 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7345)
  AUTHORS   Chatterjee A, Upadhyay S, Chang X, Nagpal JK, Trink B, Sidransky D
  TITLE     U-box-type ubiquitin E4 ligase, UFD2a attenuates cisplatin mediated 
            degradation of DeltaNp63alpha.
  JOURNAL   Cell Cycle. 2008 May 1. 7(9):1231-7.
  PUBMED    18418053
REFERENCE   2  (bases 1 to 7345)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Cycle.
            2008 May 1. 7(9):1231-7."
COMMENT     Sequence Label: deltaNp63alpha-FLAG
FEATURES             Location/Qualifiers
     source          1..7345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2459..2482
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2533..2550)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2539..2763
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2809..3237
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2896..2915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3106..3127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3246..3266)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3251..3580
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3431..3566
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3493..3512
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3629..4654
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3629..4654
                     /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    4784..4905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4821..4840)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4875..4894
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4954..4970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4954..4970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4967..4989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4978..4994
                     /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(5002..5008,5009..5026,5027..5032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5047..5068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5185..5202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5356..5944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5436..5455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6115..6906,6907..6975))
                     /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     6738..6757
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6976..7080)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7155..7174)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc atgttgtacc tggaaaacaa
      721 tgcccagact caatttagtg agccacagta cacgaacctg gggctcctga acagcatgga
      781 ccagcagatt cagaacggct cctcgtccac cagtccctat aacacagacc acgcgcagaa
      841 cagcgtcacg gcgccctcgc cctacgcaca gcccggctcc accttcgatg ctctctctcc
      901 atcacccgcc atcccctcca acaccgacta cccaggcccg cacagtttcg acgtgtcctt
      961 ccagcagtcg agcaccgcca agtcggccac ctggacgtat tccactgaac tgaagaaact
     1021 ctactgccaa attgcaaaga catgccccat ccagatcaag gtgatgaccc cacctcctca
     1081 gggagctgtt atccgcgcca tgcctgtcta caaaaaagct gagcacgtca cggaggtggt
     1141 gaagcggtgc cccaaccatg agctgagccg tgaattcaac gagggacaga ttgcccctcc
     1201 tagtcatttg attcgagtag aggggaacag ccatgcccag tatgtagaag atcccatcac
     1261 aggaagacag agtgtgctgg taccttatga gccaccccag gttggcactg aattcacgac
     1321 agtcttgtac aatttcatgt gtaacagcag ttgtgttgga gggatgaacc gccgtccaat
     1381 tttaatcatt gttactctgg aaaccagaga tgggcaagtc ctgggccgac gctgctttga
     1441 ggcccggatc tgtgcttgcc caggaagaga caggaaggcg gatgaagata gcatcagaaa
     1501 gcagcaagtt tcggacagta caaagaacgg tgatggtacg aagcgcccgt ttcgtcagaa
     1561 cacacatggt atccagatga catccatcaa gaaacgaaga tccccagatg atgaactgtt
     1621 atacttacca gtgaggggcc gtgagactta tgaaatgctg ttgaagatca aagagtccct
     1681 ggaactcatg cagtaccttc ctcagcacac aattgaaacg tacaggcaac agcaacagca
     1741 gcagcaccag cacttacttc agaaacagac ctcaatacag tctccatctt catatggtaa
     1801 cagctcccca cctctgaaca aaatgaacag catgaacaag ctgccttctg tgagccagct
     1861 tatcaaccct cagcagcgca acgccctcac tcctacaacc attcctgatg gcatgggagc
     1921 caacattccc atgatgggca cccacatgcc aatggctgga gacatgaatg gactcagccc
     1981 cacccaggca ctccctcccc cactctccat gccatccacc tcccactgca cacccccacc
     2041 tccgtatccc acagattgca gcattgtcag tttcttagcg aggttgggct gttcatcatg
     2101 tctggactat ttcacgaccc aggggctgac caccatctat cagattgagc attactccat
     2161 ggatgatctg gcaagtctga aaatccctga gcaatttcga catgcgatct ggaagggcat
     2221 cctggaccac cggcagctcc acgaattctc ctccccttct catctcctgc ggaccccaag
     2281 cagtgcctct acagtcagtg tgggctccag tgagacccgg ggtgagcgtg ttattgatgc
     2341 tgtgcgattc accctccgcc agaccatctc tttcccaccc cgagatgagt ggaatgactt
     2401 caactttgac atggatgctc gccgcaataa gcaacagcgc atcaaagagg agggggagga
     2461 ctacaaggac gacgatgaca agtagtaggc ggccgctcga gtctagaggg cccgtttaaa
     2521 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2581 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2641 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2701 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2761 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2821 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2881 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2941 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3001 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3061 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3121 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3181 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3241 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3301 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3361 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3421 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3481 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3541 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3601 atccattttc ggatctgatc agcacgtgat gaaaaagcct gaactcaccg cgacgtctgt
     3661 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     3721 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     3781 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     3841 gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat
     3901 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     3961 tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag
     4021 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4081 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4141 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4201 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     4261 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     4321 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     4381 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct
     4441 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4501 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4561 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4621 acgccccagc actcgtccga gggcaaagga atagcacgtg ctacgagatt tcgattccac
     4681 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4741 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4801 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4861 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4921 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4981 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5041 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5101 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5161 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5221 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5281 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5341 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5401 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5461 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5521 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5581 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5641 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5701 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5761 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5821 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5881 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5941 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6001 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6061 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6121 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6181 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6241 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6301 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6361 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6421 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6481 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6541 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6601 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6661 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6721 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6781 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6841 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6901 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6961 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7021 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7081 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7141 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7201 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7261 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     7321 gatgtacggg ccagatatac gcgtt
//
