LOCUS       sequence-320621-        4850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER targeted transmembrane domain with monomeric EGFP oriented 
            towards the cytoplasm. Used for assessing fluorescent protein 
            oligomerization..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4850)
  AUTHORS   Costantini LM, Fossati M, Francolini M, Snapp EL
  TITLE     Assessing the tendency of fluorescent proteins to oligomerize under 
            physiologic conditions.
  JOURNAL   Traffic. 2012 May;13(5):643-9. doi: 
            10.1111/j.1600-0854.2012.01336.x. Epub 2012 Feb 20.
  PUBMED    22289035
REFERENCE   2  (bases 1 to 4850)
  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"; doi: 
            "10.1111/j.1600-0854.2012.01336.x"; journalName: "Traffic"; date: 
            "2012-05"; volume: "13"; issue: "5"; pages: "643-9"
COMMENT     Sequence Label: CyTERM-mGFP
FEATURES             Location/Qualifiers
     source          1..4850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      731..740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(737..739,740..742,743..1456)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(782..803)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1043..1062)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    1579..1700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1616..1635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1670..1689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1707..2162)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1844..1865)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2056..2075
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2189..2293
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2295..2652
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2295..2486
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2318..2338)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2503..2638
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2687..3481
                     /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             2687..3481
                     /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(2741..2760)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3351..3370
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3669..3688)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3713..3760
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4089..4677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4578..4597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggactca gatctggtac cccttgcagc ttccagttgt caggaagaga aggctcccat
      601 ggatcctgtg gtggtgctgg ggctctgtct ctcctgtttg cttctccttt cactctggaa
      661 acagagctat gggggaggga agcttcgaat tctgcagtcg acggtaccgc gggcccggga
      721 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      781 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      841 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      901 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      961 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1021 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1081 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1141 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1201 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1261 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1321 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     1381 gcgcgatcac atggtcctta aggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1441 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     1501 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1561 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1621 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1681 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     1741 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     1801 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     1861 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     1921 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     1981 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2041 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2101 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2161 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2221 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2281 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     2341 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2401 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2461 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2521 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2581 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2641 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2701 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2761 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2821 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     2881 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2941 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3001 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3061 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3121 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3181 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     3241 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3301 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3361 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3421 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3481 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3541 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3601 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     3661 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3721 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3781 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3841 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3901 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     3961 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4021 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4081 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4141 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4201 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4261 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4321 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4381 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4441 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4501 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4561 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4621 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4681 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4741 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     4801 atagtaatca attacggggt cattagttca tagcccatat atggagttcc
//
