LOCUS       sequence-486464-        5351 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Mammalian secretory granule marker using ANF emiRFP marker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5351)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 5351)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: ANF emiRFP 670
FEATURES             Location/Qualifiers
     source          1..5351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        133..436
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        437..640
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     591..611
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1162..1171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2211..2332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2248..2267)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2302..2321
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2339..2794)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2476..2497)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2688..2707
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2821..2925
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2927..3284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2927..3118
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2950..2970)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3135..3270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3319..4113
                     /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             3319..4113
                     /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(3373..3392)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3983..4002
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4301..4320)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4345..4392
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4721..5309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5210..5229
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
       61 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      121 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      181 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      241 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      301 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      361 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      421 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      481 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      541 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      601 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      661 ccgctagcgc taccggactc agatctcgag atgggctcct tctccatcac caagggcttc
      721 ttcctcttcc tggccttttg gctcccaggc catattggag caaatcccgt atacagtgcg
      781 gtgtccaaca cagatctgat ggatttcaag aacctgctag accacctgga ggagaagatg
      841 ccggtagaag atgaggtcat gcctccgcag gccctgagcg agcagaccga tgaagcgggg
      901 gcggcactta gctccctctc tgaggtgcct ccctggactg gggaagtcaa cccgtctcag
      961 agagatggag gtgctctcgg gcgcggcccc tgggacccct ccgatagatc tgccctcttg
     1021 aaaagcaaac tgagggctct gctcgctggc cctcggagcc tgcgaaggtc aagctgcttc
     1081 gggggtagga ttgacaggat tggagcccag agcggactag gctgcaacag cttccggtac
     1141 cgtcgtggcg gcggaccggt cgccaccatg gcggaaggct ccgtcgccag gcagcctgac
     1201 ctcttgacct gcgaacatga agagatccac ctcgccggct cgatccagcc gcatggcgcg
     1261 cttctggtcg tcagcgaaca tgatcatcgc gtcatccagg ccagcgccaa cgccgcggaa
     1321 tttctgaatc tcggaagcgt actcggcgtt ccgctcgccg agatcgacgg cgatctgttg
     1381 atcaagatcc tgccgcatct cgatcccacc gccgaaggca tgccggtcgc ggtgcgctgc
     1441 cggatcggca atccctctac ggagtactgc ggtctgatgc atcggcctcc ggaaggcggg
     1501 ctgatcatcg aactcgaacg tgccggcccg tcgatcgatc tgtcaggcac gctggcgccg
     1561 gcgctggagc ggatccgcac ggcgggttca ctgcgcgcgc tgtgcgatga caccgtgctg
     1621 ctgtttcagc agtgcaccgg ctacgaccgg gtgatggtgt atcgtttcga tgagcaaggc
     1681 cacggcctgg tattctccga gtgccatgtg cctgggctcg aatcctattt cggcaaccgc
     1741 tatccgtcgt cgactgtccc gcagatggcg cggcagctgt acgtgcggca gcgcgtccgc
     1801 gtgctggtcg acgtcaccta tcagccggtg ccgctggagc cgcggctgtc gccgctgacc
     1861 gggcgcgatc tcgacatgtc gggctgcttc ctgcgctcga tgtcgccgtg ccatctgcag
     1921 ttcctgaagg acatgggcgt gcgcgccacc ctggcggtgt cgctggtggt cggcggcaag
     1981 ctgtggggcc tggttgtctg tcaccattat ctgccgcgct tcatccgttt cgagctgcgg
     2041 gcgatctgca aacggctcgc cgaaaggatc gcgacgcgga tcaccgcgct tgagagctaa
     2101 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2161 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2221 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2281 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     2341 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2401 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2461 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2521 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2581 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2641 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2701 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2761 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2821 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2881 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     2941 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3061 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3121 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3181 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3241 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3301 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3361 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3421 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3481 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     3541 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3601 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3661 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3721 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3781 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3841 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3901 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3961 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4021 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4081 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4141 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4201 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4261 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4321 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4381 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4441 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4501 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     4561 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4621 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4681 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4741 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4801 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4861 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4921 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4981 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5041 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5101 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5161 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5221 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5281 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5341 tggccttttg c
//
