LOCUS       Exported                6712 bp ds-DNA     circular SYN 09-JUN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6712)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 6, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6712
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    5..61
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    185..306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(313..768)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        795..899
                     /gene="bla"
                     /label=AmpR promoter
     promoter        901..1258
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        901..1092
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      1109..1244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1293..2087
                     /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             1293..2087
                     /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"
     polyA_signal    2319..2366
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2695..3283
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     enhancer        3458..3761
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3762..3965
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3998..4007
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4004..4693
                     /codon_start=1
                     /label=mScarlet-I
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDEL"
     CDS             4739..5263
                     /codon_start=1
                     /label=RBD
                     /translation="MSAPLGPRGRPAPTPAASQPPPQPEMPDLSHLTEEERKIILAVMD
                     RQKKEEEKEQSVLKKLHQQFEMYKEQVKKMGEESQQQQEQKGDAPTCGICHKTKFADGC
                     GHNCSYCQTKFCARCGGRVSLRSNKVMWVCNLCRKQQEILTKSGAWFYNSGSNTLQQPD
                     QKVPRGLRNEEA"
     CDS             5309..6004
                     /codon_start=1
                     /label=mNeongreen
                     /translation="SKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEEL
                     NLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDGA
                     SLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIISTF
                     KWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQKA
                     FTDVMGMDEL"
     regulatory      5558..5567
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6050..6712
                     /label=Rab3aQ81L
ORIGIN
        1 actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc
       61 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
      121 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
      181 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
      241 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
      301 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
      361 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
      421 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
      481 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
      541 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
      601 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
      661 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
      721 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
      781 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      841 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
      901 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      961 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     1021 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1081 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     1141 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     1201 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     1261 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     1321 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     1381 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     1441 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     1501 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     1561 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     1621 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     1681 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     1741 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     1801 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     1861 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     1921 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     1981 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     2041 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     2101 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     2161 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     2221 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     2281 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     2341 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     2401 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     2461 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     2521 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     2581 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2641 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2701 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2761 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2821 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2881 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2941 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3001 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3061 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3121 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3181 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3241 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3301 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3361 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     3421 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     3481 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     3541 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     3601 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     3661 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     3721 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     3781 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     3841 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     3901 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     3961 gagctggttt agtgaaccgt cagatccgct agcgctagcc accatggtga gcaagggcga
     4021 ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg gagggctcca tgaacggcca
     4081 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     4141 gctgaaggtg accaagggtg gccccctgcc cttctcctgg gacatcctgt cccctcagtt
     4201 catgtacggc tccagggcct tcatcaagca ccccgccgac atccccgact actataagca
     4261 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgccgt
     4321 gaccgtgacc caggacacct ccctggagga cggcaccctg atctacaagg tgaagctccg
     4381 cggcaccaac ttccctcctg acggccccgt aatgcagaag aagacaatgg gctgggaagc
     4441 gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag ggcgacatta agatggccct
     4501 gcgcctgaag gacggcggcc gctacctggc ggacttcaag accacctaca aggccaagaa
     4561 gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag ttggacatca cctcccacaa
     4621 cgaggactac accgtggtgg aacagtacga acgctccgag ggccgccact ccaccggcgg
     4681 catggacgag ctgggcggag gcggtggttc agggggaggg ggatctggag gtggtggcat
     4741 gtctgctccg ttaggtcccc gtggtcgtcc ggctcccact ccggctgcat ctcaacctcc
     4801 tccgcagccc gagatgccgg acctcagcca cctcacggaa gaggagagga aaatcatcct
     4861 ggctgtcatg gatcgtcaga agaaagaaga ggagaaggag cagtccgtgc tcaagaagct
     4921 gcaccaacaa tttgaaatgt ataaggagca agtcaagaag atgggagagg aatcgcagca
     4981 gcagcaagag cagaagggtg atgccccgac ctgtggcatc tgccacaaga caaaatttgc
     5041 agatggatgc ggccataatt gttcctattg ccaaaccaag ttctgtgctc gatgtggagg
     5101 tcgagtgtct ttacgctcaa acaaggttat gtgggtgtgt aatttgtgcc gaaaacaaca
     5161 agaaatcctc actaaatcag gagcatggtt ttataatagt gggtctaaca cactgcagca
     5221 acctgatcaa aaggttcctc gagggcttcg aaatgaggaa gccggcggag gcggtggttc
     5281 agggggaggg ggatctggag gtggtggcag caagggcgag gaggataaca tggcctctct
     5341 cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt
     5401 gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa
     5461 gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca
     5521 gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg
     5581 ataccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg
     5641 ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc
     5701 tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa
     5761 gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa
     5821 tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc
     5881 taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa
     5941 gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga
     6001 gctgggcgga ggcggtggtt cagggggagg gggatctgga ggtggtggca tggcctcagc
     6061 cacagactct cgatatgggc agaaggagtc ctcagaccag aacttcgact atatgttcaa
     6121 gatcctgatc attggtaaca gcagtgtggg caaaacctca ttccttttcc gctacgcaga
     6181 cgactctttt actccagcct ttgtcagcac tgtgggcata gacttcaagg tcaaaaccat
     6241 ctaccgcaat gacaagagga tcaagctgca gatctgggac acagcaggac tggagcggta
     6301 ccgaaccatc accacagcct actaccgggg cgccatgggc ttcattctaa tgtatgacat
     6361 caccaatgag gagtccttca atgcagtgca ggactggtcc actcagatca aaacttactc
     6421 atgggacaat gcccaggtgc tgctggtggg gaacaagtgc gacatggagg acgagcgagt
     6481 ggtgtcctca gaacgaggcc ggcagctggc cgaccacctg ggctttgagt tctttgaggc
     6541 cagcgccaag gacaacatta atgtcaagca gacctttgaa cgtctggtgg acgtgatctg
     6601 tgagaagatg tcggagtccc tagatactgc agaccctgca gtcacgggtg ccaagcaggg
     6661 cccacagctc accgaccagc aggcaccacc tcatcaggat tgcgcctgct aa
//
