LOCUS       sequence-220415-        6254 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 6254)
  AUTHORS   Presley JF, Cole NB, Schroer TA, Hirschberg K, Zaal KJ, 
            Lippincott-Schwartz J
  TITLE     ER-to-Golgi transport visualized in living cells.
  JOURNAL   Nature. 1997 Sep 4. 389(6646):81-5.
  PUBMED    9288971
REFERENCE   2  (bases 1 to 6254)
  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: "Nature. 
            1997 Sep 4. 389(6646):81-5."
COMMENT     Sequence Label: pEGFP-VSVG
FEATURES             Location/Qualifiers
     source          1..6254
                     /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(625..672,673..2157)
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein, 
                     thermosensitive tsO45 mutant"
                     /label=VSV-G tsO45
                     /note="thermosensitive variant of VSV-G Orsay"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAAIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNDICPTVHNSTTWHSDYKVKGLCDSNLISMDITSFSEDGELSSLGKEGTGFR
                     SNYFAYETGDKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPVFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDETLFFFGDTGLSKNPIEFVEGW
                     FSSWKSSIASFFFIIGLIIGLFLVLRVGIYLCIKLKHTKKRQIYTDIEMNRLG"
     CDS             join(2200..2202,2203..2205,2206..2919)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2245..2266)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2506..2525)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2853..2874
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3042..3163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3079..3098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3133..3152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3170..3625)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3307..3328)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3519..3538
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3652..3756
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3758..4115
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3758..3949
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3781..3801)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3966..4101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4150..4944
                     /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             4150..4944
                     /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(4204..4223)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4814..4833
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5132..5151)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5176..5223
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5552..6140
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6041..6060
                     /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 atctcgccgc caccatgaag tgccttttgt acttagcttt tttattcatc
      661 ggggtgaatt gcaagttcac catagttttt ccacacaacc aaaaaggaaa ctggaaaaat
      721 gttccttcca attaccatta ttgcccgtca agctcagatt taaattggca taatgactta
      781 ataggcacag ccttacaagt caaaatgccc aagagtcaca aggctattca agcagacggt
      841 tggatgtgtc atgcttccaa atgggtcact acttgtgatt tccgctggta cggaccgaag
      901 tatataacac attccatccg atccttcact ccatctgtag aacaatgcaa ggaaagcatt
      961 gaacaaacga aacaaggaac ttggctgaat ccaggcttcc ctcctcaaag ttgtggatat
     1021 gcaactgtga cggatgctga agcagcgatt gtccaggtga ctcctcacca tgtgcttgtt
     1081 gatgaataca caggagaatg ggttgattca cagttcatca acggaaaatg cagcaatgac
     1141 atatgcccca ctgtccataa ctccacaacc tggcattccg actataaggt caaagggcta
     1201 tgtgattcta acctcatttc catggacatc acctccttct cagaggacgg agagctatca
     1261 tccctaggaa aggagggcac agggttcaga agtaactact ttgcttatga aaccggagac
     1321 aaggcctgca aaatgcagta ctgcaagcat tggggagtca gactcccatc aggtgtctgg
     1381 ttcgagatgg ctgataagga tctctttgct gcagccagat tccctgaatg cccagaaggg
     1441 tcaagtatct ctgctccatc tcagacctca gtggatgtaa gtctcattca ggacgttgag
     1501 aggatcttgg attattccct ctgccaagaa acctggagca aaatcagagc gggtcttccc
     1561 atctctccag tggatctcag ctatcttgct cctaaaaacc caggaaccgg tcctgtcttt
     1621 accataatca atggtaccct aaaatacttt gagaccagat acatcagagt cgatattgct
     1681 gctccaatcc tctcaagaat ggtcggaatg atcagtggaa ctaccacaga aagggaactg
     1741 tgggatgact gggctccata tgaagacgtg gaaattggac ccaatggagt tctgaggacc
     1801 agttcaggat ataagtttcc tttatatatg attggacatg gtatgttgga ctccgatctt
     1861 catcttagct caaaggctca ggtgtttgaa catcctcaca ttcaagacgc tgcttcgcag
     1921 cttcctgatg atgagacttt attttttttt ggtgatactg ggctatccaa aaatccaatc
     1981 gagtttgtag aaggttggtt cagtagttgg aagagctcta ttgcctcttt tttctttatc
     2041 atagggttaa tcattggact attcttggtt ctccgagttg gtatttatct ttgcattaaa
     2101 ttaaagcaca ccaagaaaag acagatttat acagacatag agatgaaccg acttggaaac
     2161 agaattctgc agtcgacggt accgcgggcc cgggatccaa tggtgagcaa gggcgaggag
     2221 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2281 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2341 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2401 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2461 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2521 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2581 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2641 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2701 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2761 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     2821 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2881 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     2941 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3001 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3061 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3121 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3181 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3241 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3301 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3361 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3421 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3481 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3541 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3601 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3661 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3721 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     3781 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3841 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3961 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4021 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4081 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4141 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4201 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4261 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4321 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4381 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4441 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4501 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4561 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4621 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4681 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4741 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4801 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4861 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4921 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4981 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5041 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5101 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5161 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5221 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5281 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5341 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5401 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5461 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5521 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5581 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5641 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5701 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5761 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5821 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5881 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5941 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6001 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6061 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6121 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6181 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6241 ttaccgccat gcat
//
