LOCUS       sequence-137084-        6203 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 6203)
  AUTHORS   Liu B, Gross M, ten Hoeve J, Shuai K
  TITLE     A transcriptional corepressor of Stat1 with an essential LXXLL 
            signature motif.
  JOURNAL   Proc Natl Acad Sci U S A. 2001 Mar 13. 98(6):3203-7.
  PUBMED    11248056
REFERENCE   2  (bases 1 to 6203)
  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: "Proc Natl 
            Acad Sci U S A. 2001 Mar 13. 98(6):3203-7."
COMMENT     Sequence Label: Flag-hPIASy
FEATURES             Location/Qualifiers
     source          1..6203
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             616..639
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      656..665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2228..2850
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2427..2446)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2874..2894)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2879..3208
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3059..3194
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3246..3265)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3247..3265)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3279..3295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3279..3295)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3292..3314)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3303..3319
                     /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(3327..3333,3334..3351,3352..3357))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3372..3393
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3510..3527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3681..4268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3761..3780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4439..5230,5231..5299))
                     /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     5062..5081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5300..5404)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5431..5886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5518..5537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5728..5749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6012..6034
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6026..6043
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6027..6043
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgcca ccatggacta caaggacgac gatgacaaga attctgcagg atctaccatg
      661 gcggcggagc tggtggaggc caaaaacatg gtgatgagtt ttcgagtctc cgaccttcag
      721 atgctcctgg gtttcgtggg ccggagtaag agtggactga agcacgagct cgtcaccagg
      781 gccctccagc tggtgcagtt tgactgtagc cctgagctgt tcaagaagat caaggagctg
      841 tacgagaccc gctacgccaa gaagaactcg gagcctgccc cacagccgca ccggcccctg
      901 gaccccctga ccatgcactc cacctacgac cgggccggcg ctgtgcccag gactccgctg
      961 gcaggcccca atattgacta ccccgtgctc tacggaaagt acttaaacgg actgggacgg
     1021 ttgcccgcca agaccctcaa gccagaagtc cgcctggtga agctgccgtt ctttaatatg
     1081 ctggatgagc tgctgaagcc caccgaatta gtcccacaga acaacgagaa gcttcaggag
     1141 agcccgtgca tcttcgcatt gacgccaaga caggtggagt tgatccggaa ctccagggaa
     1201 ctgcagcccg gagttaaagc cgtgcaggtc gtcctgagaa tctgttactc agacaccagc
     1261 tgccctcagg aggaccagta cccgcccaac atcgctgtga aggtcaacca cagctactgc
     1321 tccgtcccgg gctactaccc ctccaataag cccggggtgg agcccaagag gccgtgccgc
     1381 cccatcaacc tcactcacct catgtacctg tcctcggcca ccaaccgcat cactgtcacc
     1441 tgggggaact acggcaagag ctactcggtg gccctgtacc tggtgcggca gctgacctca
     1501 tcggagctgc tgcagaggct gaagaccatt ggggtaaagc acccggagct gtgcaaggca
     1561 ctggtcaagg agaagctgcg ccttgatcct gacagcgaga tcgccaccac cggtgtgcgg
     1621 gtgtccctca tctgtccgct ggtgaagatg cggctctccg tgccctgccg ggcagagacc
     1681 tgcgcccacc tgcagtgctt cgacgccgtc ttctacctgc agatgaacga gaagaagccc
     1741 acctggatgt gccccgtgtg cgacaagcca gccccctacg accagctcat catcgacggg
     1801 ctcctctcga agatcctgag cgagtgtgag gacgccgacg agatcgagta cctggtggac
     1861 ggctcgtggt gcccgatccg cgccgaaaag gagcgcagct gcagcccgca gggcgccatc
     1921 ctcgtgctgg gcccctcgga cgccaatggg ctcctgcccg cccccagcgt caacgggagc
     1981 ggtgccctgg gcagcacggg tggcggcggc ccggtgggca gcatggagaa tgggaagccg
     2041 ggcgccgatg tggtggacct cacgctggac agctcatcgt cctcggagga tgaggaggag
     2101 gaggaagagg aggaggaaga cgaggacgaa gaggggcccc ggcccaagcg ccgctgcccc
     2161 ttccagaagg gcctggtgcc ggcctgctga ccccggccgc acactgactc gacatcgatt
     2221 aaagcttggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     2281 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     2341 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     2401 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     2461 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     2521 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     2581 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     2641 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     2701 ccttctgatt ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc
     2761 tggccatgcc caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt
     2821 tgggtccact cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt
     2881 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2941 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3001 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3061 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3121 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3181 cttttttgga ggcctaggct tttgcaaaaa gctcctcgag gaactgaaaa accagaaagt
     3241 taattcccta tagtgagtcg tattaaattc gtaatcatgt catagctgtt tcctgtgtga
     3301 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3361 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3421 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3481 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3541 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3601 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3661 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3721 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3781 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3841 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3901 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3961 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4021 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4081 gagttcttga agtgtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4321 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4381 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4441 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4501 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4561 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4621 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4681 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4741 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4801 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4861 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4921 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4981 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5041 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5101 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5161 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5221 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5281 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5341 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5401 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     5461 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5521 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5581 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5641 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5701 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5761 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5821 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     5881 caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5941 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6001 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgatctatac
     6061 attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat
     6121 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
     6181 atgtccaata tgaccgccat gtt
//
