LOCUS       sequence-155909-        6826 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 6826)
  AUTHORS   Liu B, Liao J, Rao X, Kushner SA, Chung CD, Chang DD, Shuai K
  TITLE     Inhibition of Stat1-mediated gene activation by PIAS1.
  JOURNAL   Proc Natl Acad Sci U S A. 1998 Sep 1. 95(18):10626-31.
  PUBMED    9724754
REFERENCE   2  (bases 1 to 6826)
  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: "Proc Natl 
            Acad Sci U S A. 1998 Sep 1. 95(18):10626-31."
COMMENT     Sequence Label: Flag-mPIAS1
FEATURES             Location/Qualifiers
     source          1..6826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     235..254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        235..253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(292..621)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      306..441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     606..626
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    650..1272
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1054..1073
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(3484..3507)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3507..3516
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3539..3742)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3568..3588)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3743..4122
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4283..4300)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4283..4299)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4292..4314)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4440..4895)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4577..4598)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4789..4808
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4922..5026
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5027..5095,5096..5887)
                     /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     complement(5245..5264)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6058..6645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6546..6565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6799..6816
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttaatac
      241 gactcactat agggaattaa ctttctggtt tttcagttcc tcgaggagct ttttgcaaaa
      301 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
      361 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      421 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      481 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      541 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      601 tccacaccct aactgacaca cattccacag ccaagctggc cgcgtaccca attcaacagg
      661 catctactga gtggacccaa cgcatgagag gacagtgcca agcaagcaac tcaaatgtcc
      721 caccggttgg gcatggccag gtagcctatg ctgtgtctgg acgtcctcct gctggtatag
      781 ttattttaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa
      841 tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa
      901 tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc
      961 ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga
     1021 gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca
     1081 ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat
     1141 aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag
     1201 gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca
     1261 gggatgccac ccaagcttta atcgatgtcg agtttttttt tttttttttt tagttttgtt
     1321 ctctgtacat ttcttttaat ttttttttcc ctaaaaaaaa ggaaaaaaaa aaccacgctt
     1381 gtatactttt ctaaacagga taaggtaaaa cagagcatag agttctcttc tgccaagtca
     1441 tttaatctgg gatggggtgg gagcagcagg gtgagagcgt cagtccaatg agataatgtc
     1501 tggtatgatg ccaaagatgg acgctgtgtc tgcgctgctc ctgctggcca caccatggcc
     1561 atggctctct ctcagactgt tggaagacac aaggctgctg ttgctgccgc tactgcttcc
     1621 gttggtggct ggcagagatg tgctccctcc tgcacttagc tggtcgagaa acatctgcga
     1681 ggaggtatac gggaaaaacc gggaggagtg caggaggtcc tggtcatctg agaccgccgc
     1741 tgcagctgcg gctagcaggg aggtgttgta atgctgattg tctcctgata agaaaggaaa
     1801 gaaatctaat ccttgtaagt cataaggcat aggcgtcatg tggaaggggt gcctgtagtc
     1861 ctggatgagg gaggtgttga tgtagcttgt atctacagca ggaaggcttg gggtgcggga
     1921 cacaggcgag gcttgatgag gaagacttaa aatgccttta ttacttagtg gtgacgtagg
     1981 agacagggaa ggacaggtcc tcttggcagg gggttcttcc tcctcttcat ctgacgagct
     2041 gtcaatggtt aggtcaatga cctcgacttt cttgttttta tttgaggact ggttgtggga
     2101 cgctacctga tgctccaatg tggagctcaa gcaaccatca actccattgt aggaggcagt
     2161 gacttcttga acctccttct ttgacctcat tggagcccac gagccatcct ccttaaactg
     2221 tatctcgtca cagtctgtgc agtactttag aatttccata aacaacccgt caataataag
     2281 gtgttcatat ggggccttct tatcacagac aggacaaacc catgttggtt ttttctcatt
     2341 catttgaatg taaagagttg catcaaaaca ctgaaggtgg gagcaggtaa gtgcccgaca
     2401 ggggattgtc agtcgcattt tcccaagtgg acacagcagc gaaacccgta ggctggtagt
     2461 agctatctca ctatctggat ctgcagttaa cttctcttta attaaagctc tagaatgatc
     2521 cggattcctt attccctttg ctcgtaacct ctgaagaaga actgttgagg acaactgttt
     2581 tacaagatat actgccatgg aataggttct tccaatttct gcagtccaag aaacaacaat
     2641 ggtatttggt actgtcgtgg acaatcggac aagtgaggtg atattaattg gtcggctagg
     2701 tcgctttggt tccacaccgt ttttagtagg tggaaggtaa cctggaaggc tgcaaggttt
     2761 tgtattcact tttacacaaa ggttgggtgg gaagtgatct tcttgtggac aactggtttc
     2821 tgataaacaa aaccttaatt ggacctgcac tgtgaagtca catttggtcc cagaaatatc
     2881 catggagctg ctgatctgct gcacctgttg tggtgtcaag gcaaatgcaa aacaggtttc
     2941 ccgaaagcgc tggctgttgt ctgaagctag actggtgggc ttgatcagtt catccaacag
     3001 gtcatagaat ggtagcttct gcagctttat gtccgggtgg actgggtgca gcgctgacgt
     3061 gagatgtggg agttccagtt catgtttggg tcccagaaga gaaacaggga gtagtgggga
     3121 tgatgcaggg tggccatcat aagtgagctg tggaatggtg gatggagaaa gagtcggagg
     3181 cataggactt gaatgtacgt tggggataga caagtccgca ggcgtcataa ttttctgagg
     3241 gaaccgcctc ctgtagagtt ctttaatttt catttgtaca gcaggactac agccagcctt
     3301 taacaaatgc agggcttttg taagaagttc gtgtttgcgt ccgtgcttgt tcctcccagc
     3361 gtagcccaac agtacttgga gttcagaaac tctaaggctc ataaccattt gctttagttc
     3421 cgcactgtcc gccatctccc gggaattcca gatctctaga ggtacccggg atcctgcaga
     3481 attcttgtca tcgtcgtcct tgtagtccat ggtggcaatt ctgacggttc actaaacgag
     3541 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aacggggcgg
     3601 ggttattacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     3661 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattgg
     3721 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     3781 aagtaggaaa gtcccgtaag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     3841 cattgacgtc aatagggggc ggacttggca tatgatacac ttgatgtact gccaagtggg
     3901 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     3961 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     4021 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     4081 ccgtaattga ttactattaa taactagtca ataatcaatg tcaacatggc ggtcatattg
     4141 gacatgagcc aatataaatg tacatattat gatatagata caacgtatgc aatggccaat
     4201 agccaatatt gatttatgct atataaccaa tgactaatat ggctaattgc caatattgat
     4261 tcaatgtata gatcagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4321 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4381 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     4441 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4501 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     4561 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4621 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     4681 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4741 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4801 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4861 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     4921 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4981 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5041 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5101 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5161 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5221 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5281 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5341 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5401 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5461 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5521 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5581 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5641 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5701 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5761 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5821 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5881 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5941 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6001 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6061 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6121 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6181 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc acacttcaag
     6241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6721 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6781 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
