LOCUS       sequence-405800-        7176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7176)
  AUTHORS   Hall MP, Huang S, Black DL
  TITLE     Differentiation-induced colocalization of the KH-type splicing 
            regulatory protein with polypyrimidine tract binding protein and the
            c-src pre-mRNA.
  JOURNAL   Mol Biol Cell. 2004 Feb . 15(2):774-86.
  PUBMED    14657238
REFERENCE   2  (bases 1 to 7176)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Cell. 2004 Feb . 15(2):774-86."
COMMENT     Sequence Label: pEGFPC1-6XHis-FLKSRP
FEATURES             Location/Qualifiers
     source          1..7176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1294..1311
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3904..4025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3941..3960)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3995..4014
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4032..4487)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4169..4190)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4381..4400
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4514..4618
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4620..4977
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4620..4811
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4643..4663)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4828..4963
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5012..5806
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5012..5806
                     /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(5066..5085)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5676..5695
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5994..6013)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6038..6085
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6414..7002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6903..6922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atccatgggc agccatcatc accatcatca cagcggcggc
     1321 cacatggtgt cggactacag cacgggagga cccccgcccg ggccgccgcc gcccgccggc
     1381 gggggcgggg gagccggagg cgccggggga ggccctccgc cgggcccgcc aggcgcgggg
     1441 gaccggggcg gcggcggtcc cggcggcggc ggcccgggcg gggggtcggc cgggggcccc
     1501 tctcagccac ccggcggagg cggcccggga atccgcaagg acgctttcgc cgacgccgtg
     1561 cagcgggccc gccagattgc agccaaaatt ggaggcgatg ctgccacgac agtgaataac
     1621 agcactcctg attttggttt tgggggccaa aagagacagt tggaagatgg agatcaaccg
     1681 gagagcaaga agctggcttc ccagggagac tcaatcagtt ctcaacttgg acccatccat
     1741 cctcccccaa ggacttcaat gacagaagag tacagggtcc cagacggcat ggtgggcctg
     1801 atcattggca gaggaggtga acaaattaac aaaatccaac aggattcagg ctgcaaagta
     1861 cagatttctc cagacagcgg tggcctaccc gagcgcagtg tgtccttgac aggagcccca
     1921 gaatctgtcc agaaagccaa gatgatgctg gatgacattg tgtctcgggg tcgtgggggc
     1981 cccccaggac agttccacga caacgccaac gggggccaga acggcaccgt gcaggagatc
     2041 atgatccccg cgggcaaggc cggcctggtc attggcaagg gcggggagac cattaagcag
     2101 ctgcaggaac gcgctggagt gaagatgatc ttaattcagg acggatctca gaatacgaat
     2161 gtggacaaac ctctccgcat cattggggat ccttacaaag tgcagcaagc ctgtgagatg
     2221 gtgatggaca tcctccggga acgtgaccaa ggcggctttg gggaccggaa tgagtacgga
     2281 tctcggattg gcggaggcat cgatgtgcca gtgcccaggc attctgttgg cgtggtcatt
     2341 ggccggagtg gagagatgat caagaagatc cagaatgatg ctggcgtgcg gatacagttc
     2401 aagcaagatg acgggacagg gcccgagaag attgctcata taatggggcc cccagacagg
     2461 tgcgagcacg cagcccggat catcaacgac ctcctccaga gcctcaggag tggtccccca
     2521 ggtcctccag ggggtccagg catgcccccg gggggccgag gccgaggaag aggccaaggc
     2581 aattggggtc cccctggcgg ggagatgacc ttctccatcc ccactcacaa gtgtgggctg
     2641 gtcatcggcc gaggtggcga gaatgtgaaa gccataaacc agcagacggg agccttcgta
     2701 gagatctccc ggcagctgcc acccaacggg gaccccaact tcaagttgtt catcatccgg
     2761 ggttcacccc agcagattga ccacgccaag cagcttatcg aggaaaagat cgagggtcct
     2821 ctctgcccag ttggaccagg cccaggtggc ccaggccctg ctggcccaat ggggcccttc
     2881 aatcctgggc ccttcaacca ggggccaccc ggggctcccc cacatgccgg ggggccccct
     2941 cctcaccagt acccacccca gggctggggc aatacctacc cccagtggca gccgcctgct
     3001 cctcatgacc caagcaaagc agctgcagcg gccgcggacc ccaacgccgc gtgggccgcc
     3061 tactactcac actactacca gcagcccccg ggccccgtcc ccggccccgc accggcccct
     3121 gcggccccac cggctcaggg tgagccccct cagcccccac ccaccggcca gtcggactac
     3181 actaaggcct gggaagagta ttacaaaaag atcggccagc agccccagca gcccggagcg
     3241 cccccacagc aggactacac gaaggcttgg gaggagtact acaagaagca agcgcaagtg
     3301 gccaccggag ggggtccagg agctccccca ggctcccagc cagactacag tgccgcctgg
     3361 gcggaatatt acagacagca ggccgcttac tacggacaga ccccaggtcc tggcggcccc
     3421 cagccgccgc ccacgcagca gggacagcag caggctcaat gaatcgaatg aatgtgaact
     3481 tcttcatctg tgaaaaatct tttttttttc cattttgttc tgtttggggg cttctgtttt
     3541 gtttggcgag agagcgatgg ctgccgtggg gagtactggg gagccctcgc ggcaagcagg
     3601 gtggggggga cttgggggca tgccgggccc tcactctctc gcctgttctg tgtctcacat
     3661 gctttttctt tcaaaattgg gatccttcca tgttgagcca gccagagaag atagcgagat
     3721 ctaaatctct gccannaaaa aaaaaaaaaa aaactcgacg gtaccgcggg cccgggatcc
     3781 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     3841 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3901 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3961 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4021 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     4081 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4141 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4201 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4261 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4321 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4381 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4441 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     4501 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4561 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     4621 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4681 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4741 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4801 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4861 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4921 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4981 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5041 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5101 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5161 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     5221 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5281 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5341 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5401 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5461 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5521 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5581 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5641 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5701 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5761 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5821 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5881 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5941 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     6001 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     6061 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     6121 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     6181 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     6241 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     6301 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6361 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6421 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6481 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6541 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6601 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6661 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6721 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6781 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6841 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6901 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6961 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7021 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7081 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     7141 ggggtcatta gttcatagcc catatatgga gttccg
//
