LOCUS       sequence-35916-e        5302 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5302)
  AUTHORS   Li J, Wang Y, Chiu SL, Cline HT
  TITLE     Membrane targeted horseradish peroxidase as a marker for correlative
            fluorescence and electron microscopy studies.
  JOURNAL   Front Neural Circuits. 2010 Feb 26;4:6. doi: 
            10.3389/neuro.04.006.2010. eCollection 2010.
  PUBMED    20204144
REFERENCE   2  (bases 1 to 5302)
  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: "Front 
            Neural Circuits."; date: "2010-02-26"; volume: "4:6. doi"; pages: " 
            10.3389/neuro.04.006.2010. eCollection 2010"
COMMENT     Sequence Label: CMV-EGFP-ssHRP-TM
FEATURES             Location/Qualifiers
     source          1..5302
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    130..264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(354..373)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(355..373)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(383..400)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(383..399)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(392..414)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      541..996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(628..647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     838..859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1022..1126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1127..1195,1196..1987)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1345..1364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2158..2746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2647..2666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2900..2917
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3034..3055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3070..3075,3076..3093,3094..3100)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3108..3124
                     /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)."
     primer_bind     3113..3135
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3132..3148
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3132..3148
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3167..3187
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3169..3187
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3288..3409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(3437..4150,4151..4153,4154..4156))
                     /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(join(3440..4150,4151..4153,4154..4156))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3482..3503)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3831..3850
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4090..4111
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        4318..5302
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggcctctcg agcctctaga
      121 taatacgtag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
      181 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
      241 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
      301 cagggggagg tgtgggaggt tttttaattc gcggccgcaa ggtacccaat tcgccctata
      361 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
      421 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      481 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
      541 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      601 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      661 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      721 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      781 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      841 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      901 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      961 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     1021 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1081 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1141 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1201 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1261 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1321 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1381 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1441 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1501 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1561 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1621 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1681 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1741 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1801 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1861 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1921 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1981 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2041 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2101 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2161 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2221 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2281 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2341 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2401 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2461 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2521 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2581 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2641 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2701 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2761 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2821 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2881 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     2941 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3001 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     3061 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     3121 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     3181 ctaaagggaa caaaagctgg agccctgcag gaagggccca agaattaaaa aacctcccac
     3241 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3301 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3361 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3421 tttacgtatt gctagcttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     3481 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
     3541 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     3601 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     3661 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     3721 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     3781 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     3841 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     3901 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     3961 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     4021 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     4081 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     4141 cgcccttgct caccatagca gatctgctcc gacgtcccca ggcagaatgg cggttcccta
     4201 aacgagcatt gcttatatag acctcccatt aggcacgcct accgcccatt tacgtcaatg
     4261 gaacgcccat ttgcgtcatt gcccctcccc attgacgtca atggggatgg gcgcgccgac
     4321 catagccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac
     4381 ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac
     4441 cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacttggca
     4501 ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc
     4561 cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct
     4621 atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg
     4681 gccaggttca atactatgta ttggccctat gccatatagt attccatata tgggttttcc
     4741 tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta
     4801 ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat
     4861 tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac
     4921 ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg
     4981 tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg
     5041 acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca
     5101 ttactattgg aaggacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta
     5161 aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac
     5221 gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat
     5281 aagcaatgct cgtttaggga ac
//
