LOCUS       sequence-133145-        4962 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 4962)
  AUTHORS   Liu Y, Min W
  TITLE     Thioredoxin promotes ASK1 ubiquitination and degradation to inhibit 
            ASK1-mediated apoptosis in a redox activity-independent manner.
  JOURNAL   Circ Res. 2002 Jun 28. 90(12):1259-66.
  PUBMED    12089063
REFERENCE   2  (bases 1 to 4962)
  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: "Circ Res. 
            2002 Jun 28. 90(12):1259-66."
COMMENT     Sequence Label: TRX1 flag
FEATURES             Location/Qualifiers
     source          1..4962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1933..2388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2020..2039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2230..2251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2514..2536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2528..2545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2706..3085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3086..3289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3240..3260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3323..3346
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3712..4334
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3911..3930)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4358..4378)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4523..4657
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4709..4728)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4710..4728)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4742..4758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4742..4758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4755..4777)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4766..4782
                     /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(4790..4796,4797..4814,4815..4820))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4835..4856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1981 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2221 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2281 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2341 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     2581 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     2641 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     2701 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     2761 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2821 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2881 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2941 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     3001 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     3061 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     3121 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3181 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     3241 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     3301 ccgtcagaat tgatctacca tggactacaa agacgatgac gacaagcttg cggccgcgaa
     3361 ttctgcagtc gacggtacca tggtgaagca gatcgagagc aagactgctt ttcaggaagc
     3421 cttggacgct gcaggtgata aacttgtagt agttgacttc tcagccacgt ggtgtgggcc
     3481 ttgcaaaatg atcaagcctt tctttcattc cctctctgaa aagtattcca acgtgatatt
     3541 ccttgaagta gatgtggatg actgtcagga tgttgcttca gagtgtgaag tcaaatgcat
     3601 gccaacattc cagtttttta agaagggaca aaaggtgggt gaattttctg gagccaataa
     3661 ggaaaagctt gaagccacca ttaatgaatt agtctaagtc tagaggatcc cgggtggcat
     3721 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3781 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3841 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3901 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3961 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     4021 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     4081 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     4141 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa
     4201 taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg
     4261 gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag
     4321 atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt
     4381 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4441 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4501 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccagttccgc ccattctccg
     4561 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4621 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcctc
     4681 gaggaactga aaaaccagaa agttaattcc ctatagtgag tcgtattaaa ttcgtaatca
     4741 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4801 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4861 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4921 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ct
//
