LOCUS       Exported                6796 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pHCMV-AmphoEnv
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6796)
  AUTHORS   Sena-Esteves M, Tebbets JC, Steffens S, Crombleholme T, Flake AW
  TITLE     Optimized large-scale production of high titer lentivirus vector 
            pseudotypes.
  JOURNAL   J Virol Methods. 2004 Dec 15. 122(2):131-9.
  PUBMED    15542136
REFERENCE   2  (bases 1 to 6796)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Virol 
            Methods. 2004 Dec 15. 122(2):131-9."
FEATURES             Location/Qualifiers
     source          1..6796
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          792..1364
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1441..3405
                     /codon_start=1
                     /gene="env"
                     /product="murine leukemia virus gPr80 envelope polyprotein 
                     precursor"
                     /label=env
                     /note="proteolytically cleaved into gp70SU and p15TM 
                     subunits"
                     /translation="MARSTLSKPPQDKINPWKPLIVMGVLLGVGMAESPHQVFNVTWRV
                     TNLMTGRTANATSLLGTVQDAFPKLYFDLCDLVGEEWDPSDQEPYVGYGCKYPAGRQRT
                     RTFDFYVCPGHTVKSGCGGPGEGYCGKWGCETTGQAYWKPTSSWDLISLKRGNTPWDTG
                     CSKVACGPCYDLSKVSNSFQGATRGGRCNPLVLEFTDAGKKANWDGPKSWGLRLYRTGT
                     DPITMFSLTRQVLNVGPRVPIGPNPVLPDQRLPSSPIEIVPAPQPPSPLNTSYPPSTTS
                     TPSTSPTSPSVPQPPPGTGDRLLALVKGAYQALNLTNPDKTQECWLCLVSGPPYYEGVA
                     VVGTYTNHSTAPANCTATSQHKLTLSEVTGQGLCMGAVPKTHQALCNTTQSAGSGSYYL
                     AAPAGTMWACSTGLTPCLSTTVLNLTTDYCVLVELWPRVIYHSPDYMYGQLEQRTKYKR
                     EPVSLTLALLLGGLTMGGIAAGIGTGTTALIKTQQFEQLHAAIQTDLNEVEKSITNLEK
                     SLTSLSEVVLQNRRGLDLLFLKEGGLCAALKEECCFYADHTGLVRDSMAKLRERLNQRQ
                     KLFETGQGWFEGLFNRSPWFTTLISTIMGPLIVLLLILLFGPCILNRLVQFVKDRISVV
                     QALVLTQQYHQLKPIEYEP"
     RBS             2436..2444
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(3500..3519)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3565..3620
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3619..3638)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3978..3994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3978..3994)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3991..4013)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4002..4018
                     /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(4026..4056)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4071..4092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4209..4226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4380..4968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4460..4479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5437..5816)
                     /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     5763..5782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6001..6105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6131..6586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6268..6289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6480..6499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6713..6735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6727..6744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6728..6744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6754..6773
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6754..6772
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"