LOCUS       sequence-456509-        6838 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 6838)
  AUTHORS   Sax JK, El-Deiry WS
  TITLE     Identification and characterization of the cytoplasmic protein TRAF4
            as a p53-regulated proapoptotic gene.
  JOURNAL   J Biol Chem. 2003 Sep 19. 278(38):36435-44.
  PUBMED    12788948
REFERENCE   2  (bases 1 to 6838)
  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"; journalName: "J Biol 
            Chem. 2003 Sep 19. 278(38):36435-44."
COMMENT     Sequence Label: pcDNA3-TRAF4
FEATURES             Location/Qualifiers
     source          1..6838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     308..327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(546..650)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(725..744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        916..1295
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1296..1499
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1450..1470
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1544..1563
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1544..1562
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3072..3090)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3073..3090)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3110..3127)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3116..3340
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3386..3814
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3473..3492)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3683..3704
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3823..3843)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3828..4157
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4008..4143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4070..4089
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4224..5018
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4278..4297)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4888..4907
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5192..5313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5229..5248)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5283..5302
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5362..5378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5362..5378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5375..5397)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5386..5402
                     /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(5410..5416,5417..5434,5435..5440))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5455..5476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5593..5610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5764..6352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5844..5863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6523..6838,1..476,477..545))
                     /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"
ORIGIN
        1 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
       61 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
      121 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
      181 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
      241 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
      301 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
      361 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
      421 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
      481 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
      541 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
      601 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      661 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
      721 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
      781 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
      841 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
      901 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
      961 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     1021 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     1081 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     1141 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     1201 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     1261 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     1321 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     1381 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     1441 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     1501 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
     1561 gggagaccca agcttggtac cgagctcgga tccccaccat gcctggcttc gactacaagt
     1621 tcctggagaa gcccaagcga cggctgctgt gcccactgtg cgggaagccc atgcgcgagc
     1681 ctgtgcaggt ttccacctgc ggccaccgtt tctgcgatac ctgcctgcag gagttcctca
     1741 gtgaaggagt cttcaagtgc cctgaggacc agcttcctct ggactatgcc aagatctacc
     1801 cagacccgga gctggaagta caagtattgg gcctgcctat ccgctgcatc cacagtgagg
     1861 agggctgccg ctggagtggg ccactacgtc atctacaggg ccacctgaat acctgcagct
     1921 tcaatgtcat tccctgccct aatcgctgcc ccatgaagct gagccgccgt gatctacctg
     1981 cacacttgca gcatgactgc cccaagcggc gcctcaagtg cgagttttgt ggctgtgact
     2041 tcagtgggga ggcctatgag agccatgagg gtatgtgccc ccaggagagt gtctactgtg
     2101 agaataagtg tggtgcccgc atgatgcggc ggctgctggc ccagcatgcc acctctgagt
     2161 gccccaagcg cactcagccc tgcacctact gcactaagga gtttgtcttt gacaccatcc
     2221 agagccacca gtaccagtgc ccaaggctgc ctgttgcctg ccccaaccaa tgtggtgtgg
     2281 gcactgtggc tcgggaggac ctgccaggcc atctgaagga cagctgtaac accgccctgg
     2341 tgctctgccc attcaaagac tccggctgca agcacaggtg ccctaagctg gcaatggcac
     2401 ggcatgtgga ggagagtgtg aagccacatc tggccatgat gtgtgccctg gtgagccggc
     2461 aacggcagga gctgcaggag cttcggcgag agctggagga gctatcagtg ggcagtgatg
     2521 gcgtgctcat ctggaagatt ggcagctatg gacggcggct acaggaggcc aaggccaagc
     2581 ccaaccttga gtgcttcagc ccagccttct acacacataa gtatggttac aagctgcagg
     2641 tgtctgcatt cctcaatggc aatggcagtg gtgagggcac acacctctca ctgtacattc
     2701 gtgtgctgcc tggtgccttt gacaatctcc ttgagtggcc ctttgcccgc cgtgtcacct
     2761 tctccctgct ggatcagagc gaccctgggc tggctaaacc acagcacgtc actgagacct
     2821 tccaccccga cccaaactgg aagaatttcc agaagccagg cacgtggcgg ggctccctgg
     2881 atgagagttc tctgggcttt ggttatccca agttcatctc ccaccaggac attcgaaagc
     2941 gaaactatgt gcgggatgat gcagtcttca tccgtgctgc tgttgaactg ccccggaaga
     3001 tcctcagctg agaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
     3061 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     3121 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3181 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3241 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3301 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3361 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3421 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3481 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3541 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3601 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3661 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3721 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3781 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3841 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3901 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3961 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4021 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4081 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4141 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4201 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4261 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4321 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4381 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4441 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4501 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4561 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4621 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4681 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4741 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4801 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4861 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4921 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4981 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5041 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5101 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5161 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5221 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5281 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5341 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5401 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5461 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5521 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5581 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5641 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5701 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5761 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5821 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5881 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5941 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6001 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6061 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6181 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6361 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6421 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6481 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6541 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6601 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6661 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6721 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6781 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgct
//
