LOCUS       sequence-76121-e        6424 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 6424)
  AUTHORS   Halaban R, Svedine S, Cheng E, Smicun Y, Aron R, Hebert DN
  TITLE     Endoplasmic reticulum retention is a common defect associated with 
            tyrosinase-negative albinism.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 May 23;97(11):5889-94.
  PUBMED    10823941
REFERENCE   2  (bases 1 to 6424)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2000-05-23"; volume: "97(11)"; pages: 
            "5889-94"
COMMENT     Sequence Label: pEGFP-TYR
FEATURES             Location/Qualifiers
     source          1..6424
                     /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"
     regulatory      2364..2373
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2370..2372,2373..2375,2376..3089)
                     /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(2370..2372,2373..2375,2376..3086)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2415..2436)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2676..2695)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3023..3044
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3212..3333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3249..3268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3303..3322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3340..3795)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3477..3498)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3689..3708
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3822..3926
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3928..4285
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3928..4119
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3951..3971)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4136..4271
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4320..5114
                     /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             4320..5114
                     /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(4374..4393)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4984..5003
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5302..5321)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5346..5393
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5722..6310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6211..6230
                     /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 attccgggaa gaatgctcct ggctgttttg
      661 tactgcctgc tgtggagttt ccagacctcc gctggccatt tccctagagc ctgtgtctcc
      721 tctaagaacc tgatggagaa ggaatgctgt ccaccgtgga gcggggacag gagtccctgt
      781 ggccagcttt caggcagagg ttcctgtcag aatatccttc tgtccaatgc accacttggg
      841 cctcaatttc ccttcacagg ggtggatgac cgggagtcgt ggccttccgt cttttataat
      901 aggacctgcc agtgctctgg caacttcatg ggattcaact gtggaaactg caagtttggc
      961 ttttggggac caaactgcac agagagacga ctcttggtga gaagaaacat cttcgatttg
     1021 agtgccccag agaaggacaa attttttgcc tacctcactt tagcaaagca taccatcagc
     1081 tcagactatg tcatccccat agggacctat ggccaaatga aaaatggatc aacacccatg
     1141 tttaacgaca tcaatattta tgacctcttt gtctggatcc attattatgt gtcaatggat
     1201 gcactgcttg ggggatatga aatctggaga gacattgatt ttgcccatga agcaccagct
     1261 tttctgcctt ggcatagact cttcttgttg cggtgggaac aagaaatcca gaagctgaca
     1321 ggagatgaaa acttcactat tccatattgg gactggcggg atgcagaaaa gtgtgacatt
     1381 tgcacagatg agtacatggg aggtcagcac cccacaaatc ctaacttact cagcccagca
     1441 tcattcttct cctcttggca gattgtctgt agccgattgg aggagtacaa cagccatcag
     1501 tctttatgca atggaacgcc cgagggacct ttacggcgta atcctggaaa ccatgacaaa
     1561 tccagaaccc caaggctccc ctcttcagct gatgtagaat tttgcctgag tttgacccaa
     1621 tatgaatctg gttccatgga taaagctgcc aatttcagct ttagaaatac actggaagga
     1681 tttgctagtc cacttactgg gatagcggat gcctctcaaa gcagcatgca caatgccttg
     1741 cacatctata tgaatggaac aatgtcccag gtacagggat ctgccaacga tcctatcttc
     1801 cttcttcacc atgcatttgt tgacagtatt tttgagcagt ggctccgaag gcaccgtcct
     1861 cttcaagaag tttatccaga agccaatgca cccattggac ataaccggga atcctacatg
     1921 gttcctttta taccactgta cagaaatggt gatttcttta tttcatccaa agatctgggc
     1981 tatgactata gctatctaca agattcagac ccagactctt ttcaagacta cattaagtcc
     2041 tatttggaac aagcgagtcg gatctggtca tggctccttg gggcggcgat ggtaggggcc
     2101 gtcctcactg ccctgctggc agggcttgtg agcttgctgt gtcgtcacaa gagaaagcag
     2161 cttcctgaag aaaagcagcc actcctcatg gagaaagagg attaccacag cttgtatcag
     2221 agccatttat aaaaggctta ggcaatagag tagggccaaa aagcctgacc tcactctaac
     2281 tcaaagtaat gtccaggttc ccagagaata tctgctggta ttttctgcag tcgacggtac
     2341 cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     2401 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2461 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2521 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2581 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2641 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2701 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2761 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2821 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     2881 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2941 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3001 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3061 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     3121 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3181 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3301 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     3361 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3421 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3481 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3541 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3601 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3661 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3721 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3781 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3841 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3901 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3961 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4021 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4081 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4141 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4201 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4261 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     4321 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4381 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4441 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4501 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4561 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4621 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4681 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4741 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4801 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4861 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4921 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4981 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5041 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5101 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5161 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5221 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5281 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5341 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     5401 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5461 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5521 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5581 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5641 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5701 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5761 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5821 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5881 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5941 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6001 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6061 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6121 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6181 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6241 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6301 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6361 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6421 gcat
//
