LOCUS       Exported                5956 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLNHX_cHS4_650, complete sequence.
ACCESSION   JN000001
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5956)
  AUTHORS   Suttiprapa S, Rinaldi G, Brindley PJ.
  TITLE     Prototypic chromatin insulator cHS4 protects retroviral transgene 
            from silencing in Schistosoma mansoni
  JOURNAL   Transgenic Res. 21 (3), 555-566 (2012)
  PUBMED    21918820
REFERENCE   2  (bases 1 to 5956)
  AUTHORS   Suttiprapa S, Rinaldi G, Brindley PJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAY-2011) Microbiology, Immunology and Tropical 
            Medicine, George Washington University, 2300 Eye Street, NW, 
            Washington, DC 20037, USA
REFERENCE   3  (bases 1 to 5956)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5956
                     /organism="Expression vector pLNHX_cHS4_650"
                     /mol_type="other DNA"
                     /db_xref="taxon:1087365"
     misc_feature    1..588
                     /label=5 Moloney murine sarcoma virus LTR
                     /note="5 Moloney murine sarcoma virus LTR"
     LTR             413..588
                     /label=5 LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    651..850
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     misc_feature    657..988
                     /label=packaging signal
                     /note="packaging signal"
     CDS             1067..1861
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /note="neomycin resistance protein"
                     /protein_id="AEO86687.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    2047..2052
                     /note="XhoI restriction binding site; Drosophila hsp70 
                     promoter was removed from the parental pLNHX by XhoI enzyme
                     digestion"
     misc_feature    2203..2505
                     /label=3 Moloney murine leukemia virus LTR
                     /note="3 Moloney murine leukemia virus LTR"
     misc_feature    2500..2505
                     /label=XbaI restriction binding site
                     /note="XbaI restriction binding site"
     misc_feature    2506..3154
                     /note="minimal cHS4 insulator (cHS4 = chicken 
                     hypersensitive site 4); Gallus gallus HS4 chromatin 
                     insulator, INSD U78775, was used as a template; comprises 
                     250 bp from the 5 active region of cHS4 fused to 400 bp of
                     the 3 region of cHS4; minimal insulator was chemically 
                     synthesized, sequence confirmed, and then inserted into the
                     XbaI site in U3 region of 3 LTR of pLNHX"
     misc_feature    3155..3451
                     /label=3 Moloney murine leukemia virus LTR
                     /note="3 Moloney murine leukemia virus LTR"
     misc_feature    3155..3160
                     /label=XbaI restriction binding site
                     /note="XbaI restriction binding site"
     misc_feature    3662..3802
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      3987
     rep_origin      complement(3988..4576)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4747..5607)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin-resistance protein"
                     /protein_id="AEO86688.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5608..5712)
                     /gene="bla"
                     /label=AmpR promoter