LOCUS       sequence-60981-e        6867 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 6867)
  AUTHORS   Galang CK, Garcia-Ramirez J, Solski PA, Westwick JK, Der CJ, 
            Neznanov NN, Oshima RG, Hauser CA
  TITLE     Oncogenic Neu/ErbB-2 increases ets, AP-1, and NF-kappaB-dependent 
            gene expression, and inhibiting ets activation blocks Neu-mediated 
            cellular transformation.
  JOURNAL   J Biol Chem. 1996 Apr 5;271(14):7992-8.
  PUBMED    8626480
REFERENCE   2  (bases 1 to 6867)
  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: "J Biol 
            Chem."; date: "1996-04-5"; volume: "271(14)"; pages: "7992-8"
COMMENT     Sequence Label: FN-Ets2
FEATURES             Location/Qualifiers
     source          1..6867
                     /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"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             928..948
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2419..2437)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2420..2437)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2457..2474)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2463..2687
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2733..3161
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2820..2839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3030..3051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3170..3190)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3175..3505
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3356..3491
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3418..3437
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3572..4366
                     /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(3626..3645)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4236..4255
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4540..4661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4577..4596)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4631..4650
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4710..4726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4710..4726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4723..4745)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4734..4750
                     /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(4758..4764,4765..4782,4783..4788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4803..4824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4941..4958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5112..5700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5192..5211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5871..6662,6663..6731))
                     /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     6494..6513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6732..6836)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttgccacc
      901 atggactaca aggacgacga cgataagcca aagaaaaagc gaaaggtcgg atccaccatg
      961 aatgactttg gaatcaagaa catggaccaa gtggcccctg tcgccaacag ttttcgtggg
     1021 acactcaagc gccagccagc ctttgacacc ttcgatggct ctctgtttgc tgtgctccct
     1081 tctctcagtg aagatcagac actccaagaa gtgcccacgg gcctggattc tgtctcccat
     1141 gactcggcca gctgcgagct gcctttgctc actccctgca gcaaggcagt gatgagccaa
     1201 gccttaaaag ccaccttcag tggcttccaa aaggagcaac gacgtcttgg catccccaaa
     1261 aacccctggc tgtggagcga gcagcaggtg tgccagtggc ttctctgggc caccaacgag
     1321 ttcagcctgg tgaatgtgaa cctgcaccag tttggcatga acggccagat gctgtgtaac
     1381 ctcggcaagg agcgcttcct ggagctggcg cctgactttg tgggtgacat cctctgggaa
     1441 catctagagc agatgatcaa agagaaccaa gaaaagacag aagaccaata tgaggaaaac
     1501 tctcacctca acgcggttcc tcattggatc aacagcaata cattaggctt cagcatggaa
     1561 caggctccat atggaatgca ggcaccaaac taccccaaag acaatctcct ggacagcatg
     1621 tgcccgccat cggccacgcc tgcagctctg ggctctgagc tccagatgtt gcccaagtct
     1681 cggctcaaca ccgtcaatgt caattactgt tccatcagcc aggacttccc cagcagcaac
     1741 gtgaatttgc tcaacaacaa ttctggaaaa cccaaggacc acgactctcc agagaacggt
     1801 ggggacagct tcgagagctc cgactcgctg ctgaggtcct ggaacagcca gtcgtcccta
     1861 ctggatgtac agcgggtacc ttccttcgag agctttgagg aggactgtag ccagtctctg
     1921 tgcctcagta agctgaccat gtccttcaag gactacatcc aagagaggag cgacccagtc
     1981 gagcaaggca aaccagttat tcctgcagca gtactggctg gcttcactgg aagcggacca
     2041 atccagttgt ggcagtttct tctggagcta ctctctgaca agtcctgtca atctttcatc
     2101 agctggacgg gggatggatg ggagttcaag cttgctgacc ccgatgaggt tgcccgccgg
     2161 tgggggaaga ggaaaaataa accaaagatg aactacgaga agctgagccg gggcttacgt
     2221 tactactacg acaagaacat catccacaag acttcgggca agcgctacgt gtaccgtttc
     2281 gtatgtgacc tgcagaactt gctgggcttc actccggagg aactgcatgc catcctgggc
     2341 gtccagcctg atacagaaga ctgatcgacc catccatcac actggcggcc gctcgagcat
     2401 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     2461 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2521 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2581 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2641 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2701 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2761 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2821 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2881 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2941 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3001 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3061 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     3121 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3181 cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca
     3241 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3301 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3361 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3421 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3481 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3541 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3601 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3661 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3721 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3781 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3841 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3901 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3961 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4021 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4081 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4141 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4201 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4261 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4321 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4381 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4441 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4501 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4561 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4621 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4681 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4741 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4801 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4921 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4981 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5041 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5101 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5161 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5281 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     5341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5521 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5641 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5701 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5761 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5821 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5881 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5941 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6001 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6061 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6121 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6181 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6241 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6301 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6361 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6421 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6481 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6541 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6601 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6661 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6721 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6781 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6841 tttccccgaa aagtgccacc tgacgtc
//
