LOCUS       sequence-386942-        6297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tepsin-GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  AUTHORS   Mattera R, Guardia CM, Sidhu SS, Bonifacino JS
  TITLE     Bivalent Motif-Ear Interactions Mediate the Association of the 
            Accessory Protein Tepsin with the AP-4 Adaptor Complex.
  JOURNAL   J Biol Chem. 2015 Dec 25;290(52):30736-49. doi: 
            10.1074/jbc.M115.683409. Epub 2015 Nov 5.
  PUBMED    26542808
REFERENCE   2  (bases 1 to 6297)
  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: 
            "10.1074/jbc.M115.683409"; journalName: "J Biol Chem"; date: 
            "2015-12-25- 25"; volume: "290"; issue: "52"; pages: "30736-49"
COMMENT     Sequence Label: pEGFP-N1-tepsin
FEATURES             Location/Qualifiers
     source          1..6297
                     /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      2237..2246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2243..2245,2246..2248,2249..2962)
                     /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(2288..2309)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2549..2568)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2896..2917
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3085..3206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3122..3141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3176..3195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3213..3668)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3350..3371)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3562..3581
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3695..3799
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3801..4158
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3801..3992
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3824..3844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4009..4144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4193..4987
                     /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             4193..4987
                     /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(4247..4266)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4857..4876
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5175..5194)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5219..5266
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5595..6183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6084..6103
                     /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 caatggctgc cgcgccgccg ctacgggacc gcctgagctt
      661 tctacaccgg ctcccgattc tcctgaaggg gacgtccgat gatgatgtcc cgtgtccggg
      721 ctacctgttt gaagagattg ctaaaatctc ccacgagtct ccgggcagca gccagtgcct
      781 gctggagtac ctcctgagcc gcctgcacag cagctccggc cacgggaagc tcaaggtgct
      841 gaagatcctg ctctatctgt gcagccacgg ctcctccttc ttcctgctca tcctcaaacg
      901 caactctgcc ttcatccagg aagctgcagc ttttgcaggg cccccagatc ctctgcacgg
      961 gaacagcttg taccagaagg ttcgcgcggc cgcgcaggac ttggggagca ccctgttctc
     1021 ggacaccgtg ttgccgctgg ctccctccca gcctctgggg accccgcctg ccacaggcat
     1081 gggctcccag gccaggccgc acagcaccct ccagggtttc ggctacagca aggaacacgg
     1141 ccgcacggct gtgaggcatc agcctgggca ggccggaggg ggctgggatg agctggacag
     1201 cggccccagc tctcagaatt cctcccagaa cagtgacctg agcagggtct cggactcggg
     1261 cagtcattcc ggcagcgaca gccattcagg ggccagccgg gagccgggtg acctggcaga
     1321 aagggtcgag gtggtggccc tgagtgactg tcagcaggag ttgagcttgg tgaggactgt
     1381 gactcgggga ccacgcgcct tcctgagccg cgaggaggca cagcacttca tcaaagcgtg
     1441 tggactgctc aactgtgagg ccgtgctgca gctgctgacc tgccacctgc gtgggaccag
     1501 tgaatgcacg cagctgaggg cgctgtgtgc catcgcctcc ctggggagca gcgacctcct
     1561 cccccaggag cacatcctcc tccgcacccg gccgtggctg caggagctca gcatgggcag
     1621 cccgggacct gtgaccaaca aggccaccaa gatcctgagg cactttgagg cctcctgtgg
     1681 gcagctgtcc cctgcccggg gcacctcagc tgagcctggc cccacagccg ccctcccagg
     1741 cccatctgac ctgctgaccg acgctgtgcc tctccctggg agccaggtct tcctgcagcc
     1801 tctgagttca accccggtct cgtcccggag ccctgctccc tcatctggga tgccgtccag
     1861 ccctgtgccc accccacccc cagatgcctc ccccattcca gcccccggag accccagcga
     1921 ggccgaggcc agactggcag aaagcaggcg gtggagacct gaacggatcc cagggggcac
     1981 ggacagccca aagagaggcc ccagcagctg tgcgtggagc cgcgactcct tgtttgctgg
     2041 catggagctg gtggcctgtc cccgcctggt gggggctggg gctgctgcgg gagagtcctg
     2101 tcctgatgct ccccgcgccc cccaaacatc gtcccagagg acagcagcca aagagcctcc
     2161 tggctcagag ccgtcagctt tcgcgttcct gaacgccctg cagtcgacgg taccgcgggc
     2221 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     2281 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2341 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2401 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2461 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2521 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2581 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2641 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     2701 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     2761 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2821 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     2881 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2941 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     3001 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3061 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3121 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3181 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     3241 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3301 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3361 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3421 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3481 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3541 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3601 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3661 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3721 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3781 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3841 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3901 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3961 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4021 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4081 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4141 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4201 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4261 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4321 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4381 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4441 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4501 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4561 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4621 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4681 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4741 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4801 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4861 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4921 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4981 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5041 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5101 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5161 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5221 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5281 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5341 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5401 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5461 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5521 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5581 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5641 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5701 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5761 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5821 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5881 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5941 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6001 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6061 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6121 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6181 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6241 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
