LOCUS       Exported                7969 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Stat3 Flag pRc/CMV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7969)
  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 7969)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genes Dev";
            date: "1995-04-15"; volume: "9(8)"; pages: "984-94"
FEATURES             Location/Qualifiers
     source          1..7969
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     103..120
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    237..258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        273..303
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    311..327
                     /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     316..338
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     335..351
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     335..351
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    382..516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(406..425)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     460..479
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(690..1484)
                     /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(801..820)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1411..1430
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1551..1880)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1565..1700
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1619..1638)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1865..1885
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1908..2421)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2103..2124)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2315..2334
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2467..2691
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2680..2697
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2717..2735
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2717..2734
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(2744..2767)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5175..5191)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5175..5191)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5281..5300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5282..5300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5345..5548)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5374..5394)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5549..5928
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6100..6119
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6194..6298
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6299..7159
                     /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(6517..6536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7330..7918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7819..7838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"