LOCUS       Exported                6003 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Beta-actin fused to monomeric near-infrared fluorescent protein 
            miRFP.
ACCESSION   .
VERSION     .
KEYWORDS    pmiRFP-beta-Actin-C-19
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6003)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 6003)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6003
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        44..347
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        348..551
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     502..522
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      590..599
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    2846..2967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2883..2902)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2937..2956
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2974..3429)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3111..3132)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3323..3342
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3456..3560
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3585..3605)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3629..3839
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3651..3847
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3698..3833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3760..3779
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3882..4676
                     /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(3936..3955)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4546..4565
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4864..4883)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4908..4955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5284..5872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5773..5792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
       61 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      121 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      181 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      241 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      301 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      361 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      421 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      481 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      541 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
      601 cgcaggacac gcaagtggaa gccccgactt tggaaccgcc gatccgagcg actgtgagag
      661 agaagaaatc catctggccg gatccatcca gccacacgga acactgctgg tggtctccga
      721 gcctgaccat agaatcattc aggcatctgc caacgccgct gaattcctga atctgggaag
      781 tgtgctggga gtccccctgg ctgagatcga cggggatctg ctgatcaaga ttctgcccca
      841 cctggaccct accgcagaag gaatgccagt ggccgtcagg tgccgcattg gaaacccctc
      901 cacagagtac gatggactga tgcatcgacc acctgaagga ggactgatca ttgagctgga
      961 acgggctggg ccacccattg atctgagcgg aacactggct cccgcactgg agcgaattcg
     1021 aactgcagga tccctgaggg ctctgtgcga cgatactgca ctgctgttcc agcagtgtac
     1081 cggctacgac cgagtgatgg tctatcggtt cgatgaacag gggcacggag aggtgtattc
     1141 agaaatacat gtgactggac tggagagcta ctttggcaac aggtatccta gctccctcgt
     1201 gccacagatg gcccggagac tgtacgaaag acagagggtg cgcgtcctgg tggatgtctc
     1261 ttatcagcct gtgccactgg agccaagact gagtcccctg actggcaggg acctggatat
     1321 gtcagggtgt tttctgcgct ctatgagtcc tactcacctg cagttcctga agaatatggg
     1381 cgtgcgggca acactggtgg tcagcctggt ggtcggaggc aaactgtggg ggctggtgat
     1441 ctgccaccat tatctgccac gcttcatcca ttttgagctg cgagcaattt gtgagctgct
     1501 ggccgaagct atcgcaaccc gcattacagc cctgtccgga ggcctcagat ctggcagcgg
     1561 tggaggcagc gcatccggcg gaagcggaag cgatgatgat atcgccgcgc tcgtcgtcga
     1621 caacggctcc ggcatgtgca aggccggctt cgcgggcgac gatgcccccc gggccgtctt
     1681 cccctccatc gtggggcgcc ccaggcacca gggcgtgatg gtgggcatgg gtcagaagga
     1741 ttcctatgtg ggcgacgagg cccagagcaa gagaggcatc ctcaccctga agtaccccat
     1801 cgagcacggc atcgtcacca actgggacga catggagaaa atctggcacc acaccttcta
     1861 caatgagctg cgtgtggctc ccgaggagca ccccgtgctg ctgaccgagg cccccctgaa
     1921 ccccaaggcc aaccgcgaga agatgaccca gatcatgttt gagaccttca acaccccagc
     1981 catgtacgtt gctatccagg ctgtgctatc cctgtacgcc tctggccgta ccactggcat
     2041 cgtgatggac tccggtgacg gggtcaccca cactgtgccc atctacgagg ggtatgccct
     2101 cccccatgcc atcctgcgtc tggacctggc tggccgggac ctgactgact acctcatgaa
     2161 gatcctcacc gagcgcggct acagcttcac caccacggcc gagcgggaaa tcgtgcgtga
     2221 cattaaggag aagctgtgct acgtcgccct ggacttcgag caagagatgg ccacggctgc
     2281 ttccagctcc tccctggaga agagctacga gctgcctgac ggccaggtca tcaccattgg
     2341 caatgagcgg ttccgctgcc ctgaggcact cttccagcct tccttcctgg gcatggagtc
     2401 ctgtggcatc cacgaaacta ccttcaactc catcatgaag tgtgacgtgg acatccgcaa
     2461 agacctgtac gccaacacag tgctgtctgg cggcaccacc atgtaccctg gcattgccga
     2521 caggatgcag aaggagatca ctgccctggc acccagcaca atgaagatca agatcattgc
     2581 tcctcctgag cgcaagtact ccgtgtggat cggcggctcc atcctggcct cgctgtccac
     2641 cttccagcag atgtggatca gcaagcagga gtatgacgag tccggcccct ccatcgtcca
     2701 ccgcaaatgc ttctagggat ccaccggatc tagataactg atcataatca gccataccac
     2761 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2821 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2881 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2941 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     3001 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3061 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3121 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3181 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3241 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3301 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3361 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3421 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3481 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3541 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3601 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3661 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3721 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3781 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3841 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3901 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3961 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4021 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4081 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4141 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4201 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4261 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4321 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4381 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4441 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4501 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4561 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4621 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4681 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4741 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4801 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4861 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4921 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4981 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5041 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5101 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     5161 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5221 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5281 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5341 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5401 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5461 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5521 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5581 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5641 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5701 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5761 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5821 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5881 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5941 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     6001 aat
//
