LOCUS       sequence-213048-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Microtubule, C1 Cloning Vector, Excitation:487, 
            Emission:509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  TITLE     Michael Davidson Unpublished
REFERENCE   2  (bases 1 to 6972)
  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-Enconsin-C-18
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      598..607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(604..606,607..609,610..1320)
                     /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(649..670)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(910..929)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1257..1278
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3751..3872
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3788..3807)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3842..3861
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3879..4334)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4016..4037)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4228..4247
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4361..4465
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4467..4824
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4467..4658
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4490..4510)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4675..4810
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4859..5653
                     /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             4859..5653
                     /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(4913..4932)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5523..5542
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5841..5860)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5885..5932
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6261..6849
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6750..6769
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc
      601 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      661 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      721 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      781 accctcgtga ccaccttgac ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
      841 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      901 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      961 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1021 cacaagctgg agtacaacta caacagccac aaggtctata tcaccgccga caagcagaag
     1081 aacggcatca aggtgaactt caagacccgc cacaacatcg aggacggcag cgtgcagctc
     1141 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1201 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1261 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagc tcaagcttcg aattcagggt ctggaagtgg tagcgcggag
     1381 ctaggagctg gcggcgacgg ccacaggggc ggcgacggcg cagtgcgaag cgaaacagca
     1441 cccgacagct acaaagtgca agataagaaa aatgcctcca gccgccctgc ctctgcaatt
     1501 tcaggacaaa ataacaacca ctcaggaaat aaaccagacc ctccgcctgt gttacgtgtt
     1561 gatgaccggc agcggctggc ccgggagcga cgtgaggaac gggagaaaca gctagctgca
     1621 agagaaatag tgtggttaga aagagaagag cgagccaggc agcactacga gaagcacctg
     1681 gaagagcgga agaagaggtt ggaggagcag aggcagaagg aggagcggag gagggctgct
     1741 gtggaggaga agcggaggca gagacttgag gaggacaaag aacgccacga agctgttgta
     1801 cggcgcacaa tggaaaggag ccagaagcca aaacagaagc ataaccgttg gtcgtgggga
     1861 ggctctctcc atgggagccc tagcatccac agtgcagatc cagacaggcg gtcagtttcc
     1921 accatgaatc tttcgaaata tgttgatccc gtcattagca agcggctctc ctcttcatct
     1981 gcaactttac taaattctcc agatagagct cgccgcctgc agctcagccc atgggagagc
     2041 agcgttgtta acagactcct gacgcccaca cattcgttcc tggccagaag taaaagcaca
     2101 gctgccttgt ctggagaagc agcatcttgc agccccatca tcatgcccta caaagctgca
     2161 cactctagaa attcgatgga tcgaccaaaa ctctttgtaa caccacctga gggctcttct
     2221 cgcaggagga tcattcatgg cacagcgagc tataaaaaag aaagagagag agaaaatgta
     2281 ctcttcctca catctggcac ccgaagggct gtatctccat ctaatcccaa agcaagacaa
     2341 ccagctcgct cccgactttg gcttccgtcc aagtctcttc ctcatttgcc tggcacaccc
     2401 agaccgacat cctccttgcc acccggctca gtcaaagctg ctcctgctca ggtccggccc
     2461 ccatcccccg gcaacatccg ccctgtcaag agggaagtca aagtggagcc tgagaagaaa
     2521 gatcctgaga aggaacctca gaaagttgcc aatgagccct cactaaaggg cagagcacct
     2581 ttagtgaagg tagaagaagc cacagttgaa gagcggacac ctgctgaacc agaagttggc
     2641 cctgctgctc cagccatggc cccagctcca gcctcggccc cagctccagc ctcggcccca
     2701 gctccagccc cggtccccac cccagccatg gtctcagccc cgtcatccac tgtgaatgcc
     2761 agtgcttctg ttaagacttc tgcaggcacc accgacccag aggaggccac aaggcttcta
     2821 gctgagaaga ggcggctggc ccgagagcag agagaaaagg aagaaaggga gaggagggag
     2881 caggaagagc ttgaaagaca aaagagagag gaattggctc aacgtgtggc tgaagagagg
     2941 acgactcgcc gtgaggagga gtcgcgcagg ctggaagccg agcaggcccg ggagaaggag
     3001 gagcagctgc agcggcaggc ggaggagcgg gcgctgcgcg agtgggagga ggcagagcgc
     3061 gcccagaggc agaaagaaga agaagctcgc gttcgtgaag aagcagagag ggtccggcag
     3121 gaacgagaga agcatttcca gagagaagag caagagcgcc tggagagaaa gaagcgactt
     3181 gaggagatta tgaaaagaac caggagaaca gaagctacag ataagaaaac cagtgatcag
     3241 agaaacggtg atatagccaa gggagctctc actggaggaa cagaggtgtc tgcacttcca
     3301 tgtacaacaa acgctccggg aaatggaaag ccagttggta gcccacatgt ggttacctca
     3361 caccagtcaa aagtgacagt ggagagcact cccgatttgg aaaaacaacc aaatgaaaat
     3421 ggtgtatctg ttcagaatga aaattttgaa gaaattataa acttacccat tggatctaaa
     3481 ccatccagat tagatgtcac caacagtgag agcccagaaa ttcctttgaa tccaattttg
     3541 gcctttgatg atgaagggac acttgggccc ctgcctcagg tagatggtgt tcagacacag
     3601 cagactgcag aagttatatg aggatccacc ggatctagat aactgatcat aatcagccat
     3661 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3721 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3781 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3841 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3901 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3961 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4021 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4081 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4141 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4201 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4261 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4321 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4381 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4441 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4501 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4561 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4621 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4681 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4741 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4801 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4861 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4921 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4981 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5041 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5101 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5161 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5221 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5281 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5341 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5401 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5461 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5521 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5581 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5641 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5701 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5761 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5821 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5881 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5941 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6001 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6061 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6121 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6181 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6241 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6301 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6361 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6421 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6481 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6541 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6601 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6661 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6721 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6781 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6841 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6901 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6961 cattagttat ta
//
