LOCUS       Exported                7969 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Stat3-C Flag pRc/CMV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7969)
  AUTHORS   Bromberg JF, Wrzeszczynska MH, Devgan G, Zhao Y, Pestell RG, 
            Albanese C, Darnell JE Jr
  TITLE     Stat3 as an oncogene.
  JOURNAL   Cell 1999 Aug 6;98(3):295-303.
  PUBMED    10458605
REFERENCE   2  (bases 1 to 7969)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; 
            date: "1999-08-6"; volume: "98(3)"; pages: "295-303"
FEATURES             Location/Qualifiers
     source          1..7969
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     296..315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(534..638)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(713..732)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        904..1283
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1284..1487
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1438..1458
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1532..1551
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1532..1550
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1641..1657
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1641..1657
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4065..4088
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(4097..4115)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4098..4115)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4135..4152)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4141..4365
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4411..4924
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4498..4517)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4708..4729
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4947..4967)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4952..5281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5132..5267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5194..5213
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5348..6142
                     /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(5402..5421)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6012..6031
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6316..6450
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6353..6372)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6407..6426
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6481..6497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6481..6497)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6494..6516)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6505..6521
                     /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(6529..6559)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6574..6595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6712..6729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6883..7471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6963..6982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7642..7969,1..533))
                     /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"