LOCUS       sequence-346391-        5327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nucelear marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5327)
  TITLE     Gadella lab plasmids
REFERENCE   2  (bases 1 to 5327)
  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: miRFP670 - H2A
FEATURES             Location/Qualifiers
     source          1..5327
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1557
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    2121..2242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2158..2177)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2212..2231
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2249..2704)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2386..2407)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2598..2617
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2731..2835
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2837..3194
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2837..3028
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2860..2880)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3045..3180
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3229..4023
                     /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             3229..4023
                     /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(3283..3302)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3893..3912
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4211..4230)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4255..4302
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4625..5213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5114..5133
                     /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 ccggtcgcca ccatggtagc aggtcatgcc tctggcagcc ccgcattcgg gaccgcctct
      661 cattcgaatt gcgaacatga agagatccac ctcgccggct cgatccagcc gcatggcgcg
      721 cttctggtcg tcagcgaaca tgatcatcgc gtcatccagg ccagcgccaa cgccgcggaa
      781 tttctgaatc tcggaagcgt actcggcgtt ccgctcgccg agatcgacgg cgatctgttg
      841 atcaagatcc tgccgcatct cgatcccacc gccgaaggca tgccggtcgc ggtgcgctgc
      901 cggatcggca atccctctac ggagtactgc ggtctgatgc atcggcctcc ggaaggcggg
      961 ctgatcatcg aactcgaacg tgccggcccg tcgatcgatc tgtcaggcac gctggcgccg
     1021 gcgctggagc ggatccgcac ggcgggttca ctgcgcgcgc tgtgcgatga caccgtgctg
     1081 ctgtttcagc agtgcaccgg ctacgaccgg gtgatggtgt atcgtttcga tgagcaaggc
     1141 cacggcctgg tattctccga gtgccatgtg cctgggctcg aatcctattt cggcaaccgc
     1201 tatccgtcgt cgactgtccc gcagatggcg cggcagctgt acgtgcggca gcgcgtccgc
     1261 gtgctggtcg acgtcaccta tcagccggtg ccgctggagc cgcggctgtc gccgctgacc
     1321 gggcgcgatc tcgacatgtc gggctgcttc ctgcgctcga tgtcgccgtg ccatctgcag
     1381 ttcctgaagg acatgggcgt gcgcgccacc ctggcggtgt cgctggtggt cggcggcaag
     1441 ctgtggggcc tggttgtctg tcaccattat ctgccgcgct tcatccgttt cgagctgcgg
     1501 gcgatctgca aacggctcgc cgaaaggatc gcgacgcgga tcaccgcgct tgagagcgtg
     1561 tacaagtact cagatctcga gctcaagctt cgaattctga tgtcgggacg cggcaagcag
     1621 ggaggcaaag ctcgcgccaa agccaagacc cgctcttctc gtgccggtct ccagttcccc
     1681 gtgggccgag tgcaccgact gctccgcaag ggcaactatg ctgagcgggt cggggccggc
     1741 gcgccggtgt acctggcggc ggtgctggag tacctgactg ccgagatcct ggagctggcg
     1801 ggcaacgccg cccgcgacaa caagaagacc cgcattatcc cgcgccactt gcagctggcc
     1861 atccgcaacg acgaggagct caacaagctg ctgggcaaag taaccatcgc tcagggtggt
     1921 gtcctgccca acatccaggc tgtgctactg cccaagaaga ccgagagtca ccacaaggcc
     1981 aaaggcaaac gggatccacc ggatctagat aactgatcat aatcagccat accacatttg
     2041 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2101 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2161 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2221 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
     2281 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2341 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     2401 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2461 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2521 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2581 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2641 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2701 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2761 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2821 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     2881 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2941 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3001 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3061 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3121 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3181 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3241 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3301 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3361 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     3421 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3481 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3541 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3601 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3661 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3721 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     3781 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3841 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3901 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3961 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4021 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4081 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4141 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     4201 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4261 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     4321 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     4381 tttcttcctt ttcccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4441 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     4501 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4561 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4621 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4681 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4741 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4801 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4861 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4921 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4981 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5041 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5101 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5161 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5221 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5281 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
