LOCUS       sequence-150296-        7812 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 7812)
  AUTHORS   Horvath CM, Wen Z, Darnell JE Jr
  TITLE     A STAT protein domain that determines DNA sequence recognition 
            suggests a novel DNA-binding domain.
  JOURNAL   Genes Dev 1995 Apr 15;9(8):984-94.
  PUBMED    7774815
REFERENCE   2  (bases 1 to 7812)
  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: "Genes Dev";
            date: "1995-04-15"; volume: "9(8)"; pages: "984-94"
COMMENT     Sequence Label: Stat1 alpha Flag pRc/CMV
FEATURES             Location/Qualifiers
     source          1..7812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3235..3258
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3271..3289)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3272..3289)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3309..3326)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3315..3539
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3585..4098
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3672..3691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3882..3903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4121..4141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4126..4455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4306..4441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4522..5316
                     /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(4576..4595)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5186..5205
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5490..5624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5527..5546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5581..5600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5655..5671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5655..5671)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5668..5690)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5679..5695
                     /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(5703..5709,5710..5727,5728..5733))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5748..5769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5886..5903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6057..6645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6137..6156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6816..7607,7608..7676))
                     /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     7439..7458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7677..7781)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat atccatcaca
      961 ctggcggccg cacaaggtgg caggatgtct cagtggtacg aacttcagca gcttgactca
     1021 aaattcctgg agcaggttca ccagctttat gatgacagtt ttcccatgga aatcagacag
     1081 tacctggcac agtggttaga aaagcaagac tgggagcacg ctgccaatga tgtttcattt
     1141 gccaccatcc gttttcatga cctcctgtca cagctggatg atcaatatag tcgcttttct
     1201 ttggagaata acttcttgct acagcataac ataaggaaaa gcaagcgtaa tcttcaggat
     1261 aattttcagg aagacccaat ccagatgtct atgatcattt acagctgtct gaaggaagaa
     1321 aggaaaattc tggaaaacgc ccagagattt aatcaggctc agtcggggaa tattcagagc
     1381 acagtgatgt tagacaaaca gaaagagctt gacagtaaag tcagaaatgt gaaggacaag
     1441 gttatgtgta tagagcatga aatcaagagc ctggaagatt tacaagatga atatgacttc
     1501 aaatgcaaaa ccttgcagaa cagagaacac gagaccaatg gtgtggcaaa gagtgatcag
     1561 aaacaagaac agctgttact caagaagatg tatttaatgc ttgacaataa gagaaaggaa
     1621 gtagttcaca aaataataga gttgctgaat gtcactgaac ttacccagaa tgccctgatt
     1681 aatgatgaac tagtggagtg gaagcggaga cagcagagcg cctgtattgg ggggccgccc
     1741 aatgcttgct tggatcagct gcagaactgg ttcactatag ttgcggagag tctgcagcaa
     1801 gttcggcagc agcttaaaaa gttggaggaa ttggaacaga aatacaccta cgaacatgac
     1861 cctatcacaa aaaacaaaca agtgttatgg gaccgcacct tcagtctttt ccagcagctc
     1921 attcagagct cgtttgtggt ggaaagacag ccctgcatgc caacgcaccc tcagaggccg
     1981 ctggtcttga agacaggggt ccagttcact gtgaagttga gactgttggt gaaattgcaa
     2041 gagctgaatt ataatttgaa agtcaaagtc ttatttgata aagatgtgaa tgagagaaat
     2101 acagtaaaag gatttaggaa gttcaacatt ttgggcacgc acacaaaagt gatgaacatg
     2161 gaggagtcca ccaatggcag tctggcggct gaatttcggc acctgcaatt gaaagaacag
     2221 aaaaatgctg gcaccagaac gaatgagggt cctctcatcg ttactgaaga gcttcactcc
     2281 cttagttttg aaacccaatt gtgccagcct ggtttggtaa ttgacctcga gacgacctct
     2341 ctgcccgttg tggtgatctc caacgtcagc cagctcccga gcggttgggc ctccatcctt
     2401 tggtacaaca tgctggtggc ggaacccagg aatctgtcct tcttcctgac tccaccatgt
     2461 gcacgatggg ctcagctttc agaagtgctg agttggcagt tttcttctgt caccaaaaga
     2521 ggtctcaatg tggaccagct gaacatgttg ggagagaagc ttcttggtcc taacgccagc
     2581 cccgatggtc tcattccgtg gacgaggttt tgtaaggaaa atataaatga taaaaatttt
     2641 cccttctggc tttggattga aagcatccta gaactcatta aaaaacacct gctccctctc
     2701 tggaatgatg ggtgcatcat gggcttcatc agcaaggagc gagagcgtgc cctgttgaag
     2761 gaccagcagc cggggacctt cctgctgcgg ttcagtgaga gctcccggga aggggccatc
     2821 acattcacat gggtggagcg gtcccagaac ggaggcgaac ctgacttcca tgcggttgaa
     2881 ccctacacga agaaagaact ttctgctgtt actttccctg acatcattcg caattacaaa
     2941 gtcatggctg ctgagaatat tcctgagaat cccctgaagt atctgtatcc aaatattgac
     3001 aaagaccatg cctttggaaa gtattactcc aggccaaagg aagcaccaga gccaatggaa
     3061 cttgatggcc ctaaaggaac tggatatatc aagactgagt tgatttctgt gtctgaagtt
     3121 cacccttcta gacttcagac cacagacaac ctgctcccca tgtctcctga ggagtttgac
     3181 gaggtgtctc ggatagtggg ctctgtagaa ttcgacagta tgatgaacac agtagactac
     3241 aaagacgatg acgataaatg agggccctat tctatagtgt cacctaaatg ctagagctcg
     3301 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3361 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3421 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3481 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3541 ttctgaggcg gaaagaacca gctggggctc gaggggggat ccccacgcgc cctgtagcgg
     3601 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3661 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3721 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3781 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3841 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3901 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3961 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4021 aatattaacg cttacaattt aaatatttgc ttatacaatc ttcctgtttt tggggctttt
     4081 ctgattatca accggggtgg gtaccgagct cgaattctgt ggaatgtgtg tcagttaggg
     4141 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4201 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4261 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4321 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4381 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4441 ctaggctttt gcaaaaagct cccgggagct tggatatcca ttttcggatc tgatcaagag
     4501 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4561 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4621 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4681 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4741 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4801 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4861 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4921 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4981 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5041 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5101 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5161 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5221 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5281 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5341 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5401 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5461 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5521 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5581 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgtcga cctcgagagc
     5641 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5701 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5761 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5821 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5881 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5941 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6901 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7141 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7441 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7741 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7801 ccacctgacg tc
//
