LOCUS       sequence-49613-e        6221 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 6221)
  AUTHORS   Koga T, Shen X, Park JS, Qiu Y, Park BC, Shyam R, Yue BY
  TITLE     Differential effects of myocilin and optineurin, two glaucoma genes,
            on neurite outgrowth.
  JOURNAL   Am J Pathol. 2010 Jan;176(1):343-52. Epub 2009 Dec 3.
  PUBMED    19959812
REFERENCE   2  (bases 1 to 6221)
  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: "Am J 
            Pathol."; date: "2010-01"; volume: "176(1)"; pages: "343-52. Epub 
            2009 Dec 3"
COMMENT     Sequence Label: pMYOCWT-EGFP
FEATURES             Location/Qualifiers
     source          1..6221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2167..2169,2170..2172,2173..2886)
                     /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             join(2167..2169,2170..2172,2173..2883)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2212..2233)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2473..2492)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2820..2841
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3009..3130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3046..3065)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3100..3119
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3137..3592)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3274..3295)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3486..3505
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3619..3723
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3725..4082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3725..3916
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3748..3768)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3933..4068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4117..4911
                     /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             4117..4911
                     /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(4171..4190)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4781..4800
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5099..5118)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5143..5190
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5519..6107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6008..6027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcccacca tggctatgag gttcttctgt
      661 gcacgttgct gcagctttgg gcctgagatg ccagctgtcc agctgctgct tctggcctgc
      721 ctggtgtggg atgtgggggc caggacagct cagctcagga aggccaatga ccagagtggc
      781 cgatgccagt ataccttcag tgtggccagt cccaatgaat ccagctgccc agagcagagc
      841 caggccatgt cagtcatcca taacttacag agagacagca gcacccaacg cttagacctg
      901 gaggccacca aagctcgact cagctccctg gagagcctcc tccaccaatt gaccttggac
      961 caggctgcca ggccccagga gacccaggag gggctgcaga gggagctggg caccctgagg
     1021 cgggagcggg accagctgga aacccaaacc agagagttgg agactgccta cagcaacctc
     1081 ctccgagaca agtcagttct ggaggaagag aagaagcgac taaggcaaga aaatgagaat
     1141 ctggccagga ggttggaaag cagcagccag gaggtagcaa ggctgagaag gggccagtgt
     1201 ccccagaccc gagacactgc tcgggctgtg ccaccaggct ccagagaagt ttctacgtgg
     1261 aatttggaca ctttggcctt ccaggaactg aagtccgagc taactgaagt tcctgcttcc
     1321 cgaattttga aggagagccc atctggctat ctcaggagtg gagagggaga caccggatgt
     1381 ggagaactag tttgggtagg agagcctctc acgctgagaa cagcagaaac aattactggc
     1441 aagtatggtg tgtggatgcg agaccccaag cccacctacc cctacaccca ggagaccacg
     1501 tggagaatcg acacagttgg cacggatgtc cgccaggttt ttgagtatga cctcatcagc
     1561 cagtttatgc agggctaccc ttctaaggtt cacatactgc ctaggccact ggaaagcacg
     1621 ggtgctgtgg tgtactcggg gagcctctat ttccagggcg ctgagtccag aactgtcata
     1681 agatatgagc tgaataccga gacagtgaag gctgagaagg aaatccctgg agctggctac
     1741 cacggacagt tcccgtattc ttggggtggc tacacggaca ttgacttggc tgtggatgaa
     1801 gcaggcctct gggtcattta cagcaccgat gaggccaaag gtgccattgt cctctccaaa
     1861 ctgaacccag agaatctgga actcgaacaa acctgggaga caaacatccg taagcagtca
     1921 gtcgccaatg ccttcatcat ctgtggcacc ttgtacaccg tcagcagcta cacctcagca
     1981 gatgctaccg tcaactttgc ttatgacaca ggcacaggta tcagcaagac cctgaccatc
     2041 ccattcaaga accgctataa gtacagcagc atgattgact acaaccccct ggagaagaag
     2101 ctctttgcct gggacaactt gaacatggtc acttatgaca tcaagctctc caagatgcgg
     2161 gatccaatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2221 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2281 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2341 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2401 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2461 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2521 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2581 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     2641 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     2701 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2761 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     2821 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2881 aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     2941 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3001 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3061 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3121 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3181 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3241 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3301 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3361 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3421 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3481 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3541 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3601 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3661 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3721 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3901 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4021 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4081 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4141 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4201 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4261 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4321 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4381 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4441 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4501 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4561 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4621 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4681 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4741 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4801 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4861 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4921 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4981 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5041 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5101 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5161 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5221 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5281 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5341 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5401 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5461 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5521 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5581 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5641 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5701 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5761 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5821 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5881 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5941 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6001 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6061 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6121 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6181 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
