LOCUS       sequence-151851-        7887 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 7887)
  AUTHORS   Wen Z, Zhong Z, Darnell JE Jr
  TITLE     Maximal activation of transcription by Stat1 and Stat3 requires both
            tyrosine and serine phosphorylation.
  JOURNAL   Cell 1995 Jul 28;82(2):241-50.
  PUBMED    7543024
REFERENCE   2  (bases 1 to 7887)
  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: "Cell"; 
            date: "1995-07-28"; volume: "82(2)"; pages: "241-50"
COMMENT     Sequence Label: Stat1 alpha Y701F pRc/CMV
FEATURES             Location/Qualifiers
     source          1..7887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        136..515
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        516..719
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     670..690
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     764..783
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        764..782
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3247..3265)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3248..3265)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3285..3302)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3291..3515
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3561..4074
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3648..3667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3858..3879
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4097..4117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4102..4431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4282..4417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4498..5292
                     /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(4552..4571)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5162..5181
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5466..5600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5503..5522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5557..5576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5631..5647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5631..5647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5644..5666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5655..5671
                     /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(5679..5685,5686..5703,5704..5709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5724..5745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5862..5879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6033..6621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6113..6132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6792..7583,7584..7652))
                     /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     7415..7434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7653..7757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7832..7851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
       61 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
      121 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
      181 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      241 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      301 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      361 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      421 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      481 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      541 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      601 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      661 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      721 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
      781 gggagaccca agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga
      841 attctgcaga tatccatcac actggcggcc gccaccatgt ctcagtggta cgaacttcag
      901 cagcttgact caaaattcct ggagcaggtt caccagcttt atgatgacag ttttcccatg
      961 gaaatcagac agtacctggc acagtggtta gaaaagcaag actgggagca cgctgccaat
     1021 gatgtttcat ttgccaccat ccgttttcat gacctcctgt cacagctgga tgatcaatat
     1081 agtcgctttt ctttggagaa taacttcttg ctacagcata acataaggaa aagcaagcgt
     1141 aatcttcagg ataattttca ggaagaccca atccagatgt ctatgatcat ttacagctgt
     1201 ctgaaggaag aaaggaaaat tctggaaaac gcccagagat ttaatcaggc tcagtcgggg
     1261 aatattcaga gcacagtgat gttagacaaa cagaaagagc ttgacagtaa agtcagaaat
     1321 gtgaaggaca aggttatgtg tatagagcat gaaatcaaga gcctggaaga tttacaagat
     1381 gaatatgact tcaaatgcaa aaccttgcag aacagagaac acgagaccaa tggtgtggca
     1441 aagagtgatc agaaacaaga acagctgtta ctcaagaaga tgtatttaat gcttgacaat
     1501 aagagaaagg aagtagttca caaaataata gagttgctga atgtcactga acttacccag
     1561 aatgccctga ttaatgatga actagtggag tggaagcgga gacagcagag cgcctgtatt
     1621 ggggggccgc ccaatgcttg cttggatcag ctgcagaact ggttcactat agttgcggag
     1681 agtctgcagc aagttcggca gcagcttaaa aagttggagg aattggaaca gaaatacacc
     1741 tacgaacatg accctatcac aaaaaacaaa caagtgttat gggaccgcac cttcagtctt
     1801 ttccagcagc tcattcagag ctcgtttgtg gtggaaagac agccctgcat gccaacgcac
     1861 cctcagaggc cgctggtctt gaagacaggg gtccagttca ctgtgaagtt gagactgttg
     1921 gtgaaattgc aagagctgaa ttataatttg aaagtcaaag tcttatttga taaagatgtg
     1981 aatgagagaa atacagtaaa aggatttagg aagttcaaca ttttgggcac gcacacaaaa
     2041 gtgatgaaca tggaggagtc caccaatggc agtctggcgg ctgaatttcg gcacctgcaa
     2101 ttgaaagaac agaaaaatgc tggcaccaga acgaatgagg gtcctctcat cgttactgaa
     2161 gagcttcact cccttagttt tgaaacccaa ttgtgccagc ctggtttggt aattgacctc
     2221 gagacgacct ctctgcccgt tgtggtgatc tccaacgtca gccagctccc gagcggttgg
     2281 gcctccatcc tttggtacaa catgctggtg gcggaaccca ggaatctgtc cttcttcctg
     2341 actccaccat gtgcacgatg ggctcagctt tcagaagtgc tgagttggca gttttcttct
     2401 gtcaccaaaa gaggtctcaa tgtggaccag ctgaacatgt tgggagagaa gcttcttggt
     2461 cctaacgcca gccccgatgg tctcattccg tggacgaggt tttgtaagga aaatataaat
     2521 gataaaaatt ttcccttctg gctttggatt gaaagcatcc tagaactcat taaaaaacac
     2581 ctgctccctc tctggaatga tgggtgcatc atgggcttca tcagcaagga gcgagagcgt
     2641 gccctgttga aggaccagca gccggggacc ttcctgctgc ggttcagtga gagctcccgg
     2701 gaaggggcca tcacattcac atgggtggag cggtcccaga acggaggcga acctgacttc
     2761 catgcggttg aaccctacac gaagaaagaa ctttctgctg ttactttccc tgacatcatt
     2821 cgcaattaca aagtcatggc tgctgagaat attcctgaga atcccctgaa gtatctgtat
     2881 ccaaatattg acaaagacca tgcctttgga aagtattact ccaggccaaa ggaagcacca
     2941 gagccaatgg aacttgatgg ccctaaagga actggattta tcaagactga gttgatttct
     3001 gtgtctgaag ttcacccttc tagacttcag accacagaca acctgctccc catgtctcct
     3061 gaggagtttg acgaggtgtc tcggatagtg ggctctgtag aattcgacag tatgatgaac
     3121 acagtataga gcatgaattt ttttcatctt ctctggcgac agttttcctt ctcatctgtg
     3181 attccctcct gctactctgt tccttcacat cctgtgtttc tagggaaatg aaagaaaggg
     3241 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     3301 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3361 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3421 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3481 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3541 gggctcgagg ggggatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3601 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3661 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3721 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3781 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3841 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3901 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3961 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaaat
     4021 atttgcttat acaatcttcc tgtttttggg gcttttctga ttatcaaccg gggtgggtac
     4081 cgagctcgaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4201 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4261 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4321 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4381 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4441 ggagcttgga tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4501 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4561 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4621 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4681 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4741 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4801 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4861 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4921 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4981 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5041 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5101 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5161 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5221 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5281 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5341 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5401 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5461 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5521 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5581 tatcttatca tgtctggatc cgtcgacctc gagagcttgg cgtaatcatg gtcatagctg
     5641 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5701 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5761 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5821 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5881 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5941 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6001 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6061 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6121 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6181 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6241 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6301 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6361 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6421 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6481 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6541 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6601 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6661 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6781 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6841 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6901 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6961 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7021 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7081 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7141 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7201 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7261 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7321 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7381 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7441 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7501 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7561 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7621 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7681 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7741 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     7801 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7861 gccagtatct gctccctgct tgtgtgt
//
