LOCUS       Exported                6550 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Fluorescent marker for hepatocytic lineage.
ACCESSION   .
VERSION     .
KEYWORDS    pALB-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6550)
  AUTHORS   Heo J, Factor VM, Uren T, Takahama Y, Lee JS, Major M, Feinstone SM,
            Thorgeirsson SS
  TITLE     Hepatic precursors derived from murine embryonic stem cells 
            contribute to regeneration of injured liver.
  JOURNAL   Hepatology. 2006 Dec;44(6):1478-86.
  PUBMED    17133486
REFERENCE   2  (bases 1 to 6550)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Hepatology."; date: "2006-12"; volume: "44(6)"; pages: "1478-86"
FEATURES             Location/Qualifiers
     source          1..6550
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(2331..2919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2411..2430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3248..3295
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3320..3339
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3527..4321)
                     /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(3638..3657)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4248..4267
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4356..4713)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4370..4505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4424..4443)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4670..4690
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4715..4819)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4846..5301
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4933..4952)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5143..5164
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5308..5429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5319..5338)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5373..5392
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5552..6271)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(5555..6271)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5597..5618)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5946..5965
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6205..6226
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      6268..6277
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     6316..6332
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6317..6333
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"