LOCUS       sequence-488282-        6112 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Secretory granule marker of endocrine cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6112)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 6112)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: mCgA EGFP
FEATURES             Location/Qualifiers
     source          1..6112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1992..2001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1998..2000,2001..2003,2004..2717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1998..2000,2001..2003,2004..2714)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2043..2064)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2304..2323)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2651..2672
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2840..2961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2877..2896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2931..2950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2968..3423)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3105..3126)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3317..3336
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3450..3554
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3556..3913
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3556..3747
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3579..3599)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3764..3899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3948..4742
                     /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             3948..4742
                     /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(4002..4021)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4612..4631
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4930..4949)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4974..5021
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5350..5938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5839..5858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attccgccac catgcgctcc accgcggttc
      601 tggcgcttct gctgtgcgcc gggcaagttt ttgcccttcc tgtgaacagc cccatgacaa
      661 aaggggacac caaggtgatg aagtgcgtcc tggaagtcat ctccgactcg ctgtccaaac
      721 ccagccccat gcctgtcagc cctgagtgtc tggagaccct ccaaggagac gagaggatcc
      781 tctctatcct gcgacaccag aatctgctga aggaacttca agacctggct cttcaaggtg
      841 ccaaggagcg ggcccagcag ccgctgaagc agcagcagcc gccgaagcag cagcagcagc
      901 agcggcagca gcagcaggag cagcaacaca gcagctttga ggatgaactc tcagaagtgt
      961 ttgagaacca gagccctgat gccaagcaca gagacgcagc agcagaagtg ccctccaggg
     1021 acactatgga gaagagaaag gattctgaca agggacaaca ggatggcttt gaggcaacca
     1081 cagagggacc taggcctcag gcctttccag agcctaacca agagtccccc atgatgggag
     1141 acagtgagtc ccctggggag gacacagcca ccaataccca atcaccaacc agcctcccca
     1201 gccaggagca tgtggaccca caggccacgg gagacagtga gagaggcctg agtgcccaac
     1261 agcaagccag aaaagccaag caagaggaga aagaagagga ggaggaagag gaggctgtgg
     1321 ctagagagaa ggctgggcct gaagaagtcc ccactgcagc atccagttcc cacttccatg
     1381 caggctacaa agcgatccag aaagatgatg gtcagtcgga ctctcaggca gtggatggag
     1441 atggaaagac agaggcttcc gaagctctgc cgtctgaagg gaagggggag ctggagcact
     1501 ctcagcagga ggaagatggg gaagaggcca tggtagggac cccccaaggt ctcttccccc
     1561 aaggcgggaa gggccgggag ctggaacata agcaggagga agaggaagag gaggaggagc
     1621 gtctgtccag agaatgggag gacaagcgat ggagcaggat ggaccagctg gccaaagagc
     1681 tgacagcaga gaagcggctg gagggcgagg atgaccctga ccgctccatg aagctctcct
     1741 tccggacccg cgcctatggc ttcagagatc ctgggcctca gctacgcagg ggctggaggc
     1801 catcttccag agaagacagt gtggaggccc gaagtgactt tgaggaaaag aaggaagagg
     1861 agggcagcgc caaccgcaga gcagaggacc aggagctaga gagcctgtca gccatcgagg
     1921 cagagctgga gaaggtggcc caccagctgc aggctttgcg gcggggaggt accgcgggcc
     1981 cgggatccat cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2041 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2101 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2161 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     2221 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2281 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2341 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2401 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     2461 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     2521 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2581 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     2641 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2701 acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     2761 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2821 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2881 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2941 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3001 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3061 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3121 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3181 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3241 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3301 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3361 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3421 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3481 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3541 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3601 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3661 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3721 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3781 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3841 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3901 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3961 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4021 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4081 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4141 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4201 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4261 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4321 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4381 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4441 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4501 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4561 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4621 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4681 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4741 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4801 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4861 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4921 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4981 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5041 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5101 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5161 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5221 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5281 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5341 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5401 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5461 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5521 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5581 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5641 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5701 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5761 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5821 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5881 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5941 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6001 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     6061 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cg
//
