LOCUS       sequence-406429-        8500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Membrane Receptor, N1 Cloning Vector, Excitation:487, 
            Emission:509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8500)
  TITLE     Michael Davidson Unpublished
REFERENCE   2  (bases 1 to 8500)
  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"
COMMENT     Sequence Label: mEmerald-ERBB2-N-18
FEATURES             Location/Qualifiers
     source          1..8500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      677..686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4499..4508
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4505..4507,4508..4510,4511..5224)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4550..4571)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4811..4830)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5158..5179
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5346..5467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5383..5402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5437..5456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5474..5929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5611..5632)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5823..5842
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5956..6060
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6062..6419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6062..6253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6085..6105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6270..6405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6454..7248
                     /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             6454..7248
                     /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(6508..6527)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7118..7137
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7436..7455)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7480..7527
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7856..8444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8345..8364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggactcagat ctcgaggcca ccatggagct ggcggccttg tgccgctggg ggctcctcct
      721 cgccctcttg ccccccggag ccgcgagcac ccaagtgtgc accggcacag acatgaagct
      781 gcggctccct gccagtcccg agacccacct ggacatgctc cgccacctct accagggctg
      841 ccaggtggtg cagggaaacc tggaactcac ctacctgccc accaatgcca gcctgtcctt
      901 cctgcaggat atccaggagg tgcagggcta cgtgctcatc gctcacaacc aagtgaggca
      961 ggtcccactg cagaggctgc ggattgtgcg aggcacccag ctctttgagg acaactatgc
     1021 cctggccgtg ctagacaatg gagacccgct gaacaatacc acccctgtca caggggcctc
     1081 cccaggaggc ctgcgggagc tgcagcttcg aagcctcaca gagatcttga aaggaggggt
     1141 cttgatccag cggaaccccc agctctgcta ccaggacacg attttgtgga aggacatctt
     1201 ccacaagaac aaccagctgg ctctcacact gatagacacc aaccgctctc gggcctgcca
     1261 cccctgttct ccgatgtgta agggctcccg ctgctgggga gagagttctg aggattgtca
     1321 gagcctgacg cgcactgtct gtgccggtgg ctgtgcccgc tgcaaggggc cactgcccac
     1381 tgactgctgc catgagcagt gtgctgccgg ctgcacgggc cccaagcact ctgactgcct
     1441 ggcctgcctc cacttcaacc acagtggcat ctgtgagctg cactgcccag ccctggtcac
     1501 ctacaacaca gacacgtttg agtccatgcc caatcccgag ggccggtata cattcggcgc
     1561 cagctgtgtg actgcctgtc cctacaacta cctttctacg gacgtgggat cctgcaccct
     1621 cgtctgcccc ctgcacaacc aagaggtgac agcagaggat ggaacacagc ggtgtgagaa
     1681 gtgcagcaag ccctgtgccc gagtgtgcta tggtctgggc atggagcact tgcgagaggt
     1741 gagggcagtt accagtgcca atatccagga gtttgctggc tgcaagaaga tctttgggag
     1801 cctggcattt ctgccggaga gctttgatgg ggacccagcc tccaacactg ccccgctcca
     1861 gccagagcag ctccaagtgt ttgagactct ggaagagatc acaggttacc tatacatctc
     1921 agcatggccg gacagcctgc ctgacctcag cgtcttccag aacctgcaag taatccgggg
     1981 acgaattctg cacaatggcg cctactcgct gaccctgcaa gggctgggca tcagctggct
     2041 ggggctgcgc tcactgaggg aactgggcag tggactggcc ctcatccacc ataacaccca
     2101 cctctgcttc gtgcacacgg tgccctggga ccagctcttt cggaacccgc accaagctct
     2161 gctccacact gccaaccggc cagaggacga gtgtgtgggc gagggcctgg cctgccacca
     2221 gctgtgcgcc cgagggcact gctggggtcc agggcccacc cagtgtgtca actgcagcca
     2281 gttccttcgg ggccaggagt gcgtggagga atgccgagta ctgcaggggc tccccaggga
     2341 gtatgtgaat gccaggcact gtttgccgtg ccaccctgag tgtcagcccc agaatggctc
     2401 agtgacctgt tttggaccgg aggctgacca gtgtgtggcc tgtgcccact ataaggaccc
     2461 tcccttctgc gtggcccgct gccccagcgg tgtgaaacct gacctctcct acatgcccat
     2521 ctggaagttt ccagatgagg agggcgcatg ccagccttgc cccatcaact gcacccactc
     2581 ctgtgtggac ctggatgaca agggctgccc cgccgagcag agagccagcc ctctgacgtc
     2641 catcatctct gcggtggttg gcattctgct ggtcgtggtc ttgggggtgg tctttgggat
     2701 cctcatcaag cgacggcagc agaagatccg gaagtacacg atgcggagac tgctgcagga
     2761 aacggagctg gtggagccgc tgacacctag cggagcgatg cccaaccagg cgcagatgcg
     2821 gatcctgaaa gagacggagc tgaggaaggt gaaggtgctt ggatctggcg cttttggcac
     2881 agtctacaag ggcatctgga tccctgatgg ggagaatgtg aaaattccag tggccatcaa
     2941 agtgttgagg gaaaacacat cccccaaagc caacaaagaa atcttagacg aagcatacgt
     3001 gatggctggt gtgggctccc catatgtctc ccgccttctg ggcatctgcc tgacatccac
     3061 ggtgcagctg gtgacacagc ttatgcccta tggctgcctc ttagaccatg tccgggaaaa
     3121 ccgcggacgc ctgggctccc aggacctgct gaactggtgt atgcagattg ccaaggggat
     3181 gagctacctg gaggatgtgc ggctcgtaca cagggacttg gccgctcgga acgtgctggt
     3241 caagagtccc aaccatgtca aaattacaga cttcgggctg gctcggctgc tggacattga
     3301 cgagacagag taccatgcag atgggggcaa ggtgcccatc aagtggatgg cgctggagtc
     3361 cattctccgc cggcggttca cccaccagag tgatgtgtgg agttatggtg tgactgtgtg
     3421 ggagctgatg acttttgggg ccaaacctta cgatgggatc ccagcccggg agatccctga
     3481 cctgctggaa aagggggagc ggctgcccca gccccccatc tgcaccattg atgtctacat
     3541 gatcatggtc aaatgttgga tgattgactc tgaatgtcgg ccaagattcc gggagttggt
     3601 gtctgaattc tcccgcatgg ccagggaccc ccagcgcttt gtggtcatcc agaatgagga
     3661 cttgggccca gccagtccct tggacagcac cttctaccgc tcactgctgg aggacgatga
     3721 catgggggac ctggtggatg ctgaggagta tctggtaccc cagcagggct tcttctgtcc
     3781 agaccctgcc ccgggcgctg ggggcatggt ccaccacagg caccgcagct catctaccag
     3841 gagtggcggt ggggacctga cactagggct ggagccctct gaagaggagg cccccaggtc
     3901 tccactggca ccctccgaag gggctggctc cgatgtattt gatggtgacc tgggaatggg
     3961 ggcagccaag gggctgcaaa gcctccccac acatgacccc agccctctac agcggtacag
     4021 tgaggacccc acagtacccc tgccctctga gactgatggc tacgttgccc ccctgacctg
     4081 cagcccccag cctgaatatg tgaaccagcc agatgttcgg ccccagcccc cttcgccccg
     4141 agagggccct ctgcctgctg cccgacctgc tggtgccact ctggaaaggc ccaagactct
     4201 ctccccaggg aagaatgggg tcgtcaaaga cgtttttgcc tttgggggtg ccgtggagaa
     4261 ccccgagtac ttgacacccc agggaggagc tgcccctcag ccccaccctc ctcctgcctt
     4321 cagcccagcc ttcgacaacc tctattactg ggaccaggac ccaccagagc ggggggctcc
     4381 acccagcacc ttcaaaggga cacctacggc agagaaccca gagtacctgg gtctggacgt
     4441 gccagtgaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc
     4501 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4561 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     4621 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     4681 caccctcgtg accaccttga cctacggcgt gcagtgcttc gcccgctacc ccgaccacat
     4741 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4801 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     4861 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     4921 gcacaagctg gagtacaact acaacagcca caaggtctat atcaccgccg acaagcagaa
     4981 gaacggcatc aaggtgaact tcaagacccg ccacaacatc gaggacggca gcgtgcagct
     5041 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     5101 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     5161 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     5221 gtaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     5281 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     5341 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5401 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5461 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     5941 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6001 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6061 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     6121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     6181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     6421 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     6481 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6541 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6601 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     6661 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6721 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6781 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6841 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6901 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6961 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     7021 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     7081 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     7141 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     7201 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     7261 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     7321 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     7381 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     7441 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     7501 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     7561 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     7621 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     7681 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     7741 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7801 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7861 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7921 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7981 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     8041 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8101 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8161 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8221 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8281 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8341 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8401 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8461 cctttttacg gttcctggcc ttttgctggc cttttgctca
//
