LOCUS       Exported                5256 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCR4-24XMS2SL-stable
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5256)
  AUTHORS   Bertrand E, Chartrand P, Schaefer M, Shenoy SM, Singer RH, Long RM
  TITLE     Localization of ASH1 mRNA particles in living yeast.
  JOURNAL   Mol Cell. 1998 Oct . 2(4):437-45.
  PUBMED    9809065
REFERENCE   2  (bases 1 to 5256)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            1998 Oct . 2(4):437-45."
FEATURES             Location/Qualifiers
     source          1..5256
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(35..53)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(74..90)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(74..90)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(87..109)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    98..114
                     /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(122..152)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    167..188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(305..322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(476..1064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(556..575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1372..1750
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5-AACGTT-3 (Sato et al., 1996)"
     primer_bind     complement(1406..1425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2296..3090)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2407..2426)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3017..3036
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(join(3439..5256,1..78))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSSELTLTKGTSPAGLNEFALRSDEPWNTGATRDRRRLKI
                     RPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPW
                     NTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARA
                     FLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRR
                     LKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSD
                     EPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTAD
                     ARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRD
                     RRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTV
                     GSKGEFAAAKFNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLYVRQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(3541..3560)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3864..3886
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3878..3895
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3879..3895
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3902..3921
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3902..3920
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"