LOCUS       sequence-414334-        6093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of dMASS_1 in mammalian cells under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6093)
  AUTHORS   Rappleye M, Wait SJ, Lee JD, Siebart JC, Torp L, Smith N, Muster J, 
            Matreyek KA, Fowler DM, Berndt A
  TITLE     Optogenetic Microwell Array Screening System: A High-Throughput 
            Engineering Platform for Genetically Encoded Fluorescent Indicators.
  JOURNAL   ACS Sens. 2023 Nov 24;8(11):4233-4244. doi: 
            10.1021/acssensors.3c01573. Epub 2023 Nov 13.
  PUBMED    37956352
REFERENCE   2  (bases 1 to 6093)
  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"; doi: 
            "10.1021/acssensors.3c01573"; journalName: "ACS Sens"; date: 
            "2023-11-24- 24"; volume: "8"; issue: "11"; pages: "4233-4244"
COMMENT     Sequence Label: pCMV-dMASS_1
FEATURES             Location/Qualifiers
     source          1..6093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     509..528
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(827..846)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    871..918
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1247..1835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1736..1755
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2010..2313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2314..2517
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2468..2488
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3622..3643
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3749..3770)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4010..4029)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    4830..4951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4867..4886)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4921..4940
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4958..5413)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5095..5116)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5307..5326
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5440..5544
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5546..5903
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5546..5737
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5569..5589)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5754..5889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5938..6093,1..639)
                     /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             join(5938..6093,1..639)
                     /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(5992..6011)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
       61 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
      121 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
      181 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
      241 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
      301 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
      361 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
      421 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
      481 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
      541 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
      601 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
      661 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
      721 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
      781 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
      841 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
      901 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
      961 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     1021 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     1081 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1141 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1201 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1261 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1321 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1381 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1441 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1501 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1561 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1621 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1681 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1741 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1801 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1861 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1921 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     1981 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2041 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2101 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2161 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2221 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2281 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     2341 gggcgtggat agtggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     2401 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     2461 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     2521 ttagtgaacc gtcagatccg ctagcgctac cggactcaga tctcgagctc aagcttaccg
     2581 gtgcgcgcca tgaagacgat catcgccctg agctacatct tctgcctggt gttcgccgac
     2641 tacaaggacg atgatgacgc catgtctggt caaccaccgc ggtctccgtc gtaaggatca
     2701 tccgcagatt ataccgcaac tacacgccac catgaggact ctgaacacct ctgccatgga
     2761 cgggactggg ctggtggtgg agagggactt ctctgttcgt atcctcactg cctgtttcct
     2821 gtcgctgctc atcctgtcca cgctcctggg gaacacgctg gtctgtgctg ccgttatcag
     2881 gttccgacac ctgcggtcca aggtgaccaa cttctttgtc atctccttgg ctgtgtcaga
     2941 tctcttggtg gccgtcctgg tcatgccctg gaaggcagtg gctgagattg ctggcttctg
     3001 gccctttggg tccttctgta acatctgggt ggcctttgac atcatgtgct ccactgcatc
     3061 catcctcaac ctctgtgtga tcagcgtgga caggtattgg gctatctcca gccctttccg
     3121 gtatgagaga aagatgaccc ccaaggcagc cttcatcctg atcagtgtgg catggacctt
     3181 gtctgtactc atctccttca tcccagtgca gctcagctgg cacaaggcaa aacccacaag
     3241 cccctctgat ggaaatgcca cttccctggc tgagaccata gacaactgtg actccagcct
     3301 cagcaggaca tatgccatct catcctctgt aatcagcttt tacatccctg tggccatcat
     3361 gattgtcacc tacaccagga tctacaggat tgctcagaaa ctgagctcag gttgtaacgt
     3421 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     3481 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     3541 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga
     3601 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3661 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga
     3721 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3781 caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa
     3841 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3901 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3961 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     4021 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4081 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaaccg
     4141 gttggaccaa ctgaaaagag aaactaaagt cctgaagact ctgtcggtga tcatgggtgt
     4201 gtttgtgtgc tgttggctac ctttcttcat cttgaactgc attttgccct tctgtgggtc
     4261 tggggagacg cagcccttct gcattgattc caacaccttt gacgtgtttg tgtggtttgg
     4321 gtgggctaat tcatccttga accccatcat ttatgccttt aatgctgatt ttcggaaggc
     4381 attttcaacc ctcttaggat gctacagact ttgccctgcg acgaataatg ccatagagac
     4441 ggtgagtatc aataacaatg gggccgcgat gttttccagc catcatgagc cacgaggctc
     4501 catctccaag gagtgcaatc tggtttacct gatcccacat gctgtgggct cctctgagga
     4561 cctgaaaaag gaggaggcag ctggcatcgc cagacccttg gagaagctgt ccccagccct
     4621 atcggtcata ttggactatg acactgacgt ctctctggag aagatccaac ccatcacaca
     4681 aaacggtcag cacccaacct gatgtacagc ggccgcgact ctagatcata atcagccata
     4741 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4801 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4861 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4921 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     4981 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5041 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5101 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5161 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5221 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5281 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5341 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5401 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5461 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5521 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     5581 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5641 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5701 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5761 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5821 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5881 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     5941 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6001 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6061 caggggcgcc cggttctttt tgtcaagacc gac
//
