LOCUS       Exported                5245 bp DNA     circular SYN 14-JAN-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5245)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 14, 2022 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    459..479
                     /locus_tag="CMV forward"
                     /label=CMV forward
                     /note="color: #c0c0c0"
     regulatory      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     misc_feature    543..989
                     /locus_tag="Tom20 plus linker"
                     /label=Tom20 plus linker
                     /note="color: #00ffff"
     misc_feature    990..1121
                     /locus_tag="CoilE"
                     /label=CoilE
                     /note="color: #80ff80"
     misc_feature    990
                     /locus_tag="AgeI site"
                     /label=AgeI site
                     /note="color: #ff9e2b"
     misc_feature    1122..1139
                     /locus_tag="Tom20 plus linker(1)"
                     /label=Tom20 plus linker
                     /label=Tom20 plus linker(1)
                     /note="color: #00ffff"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     misc_feature    1140..1850
                     /locus_tag="mCherry"
                     /label=mCherry
                     /note="color: #ff80ff"
     polyA_signal    1973..2094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(2101..2556)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2583..2687
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        2689..3046
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      2897..3032
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3081..3875
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4107..4154
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     rep_origin      4483..5071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      541 ccatggtggg tcggaacagc gccatcgccg ccggtgtatg cggggccctt ttcattgggt
      601 actgcatcta cttcgaccgc aaaagacgaa gtgaccccaa cttcaagaac aggcttcgag
      661 aacgaagaaa gaaacagaag cttgccaagg agagagctgg gctttccaag ttacctgacc
      721 ttaaagatgc tgaagctgtt cagaagttct tccttgaaga aatacagctt ggtgaagagt
      781 tactagctca aggtgaatat gagaagggcg tagaccatct gacaaatgca attgctgtgt
      841 gtggacagcc acagcagtta ctgcaggtct tacagcaaac tcttccacca ccagtgttcc
      901 agatgcttct gactaagctc ccaacaatta gtcagagaat tgtaagtgct cagagcttgg
      961 ctgaagatga tgtggaaggc ggtagcggga tgctggaaat tgaagcggcg tttctggaac
     1021 gtgagaatac cgcgctggaa acccgtgtgg cggaactgcg tcagcgtgtg cagcgtctgc
     1081 gtaatcgtgt gagccagtat cgtacccgtt atggcccgct ggatccaccg gtcgccacca
     1141 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1201 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1261 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     1321 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     1381 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     1441 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     1501 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     1561 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     1621 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     1681 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     1741 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1801 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa agcggccgcg
     1861 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1921 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     1981 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2041 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     2101 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2161 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2221 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2281 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2341 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2401 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2461 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2521 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     2581 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2641 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     2701 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2761 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2821 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2881 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2941 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3001 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3061 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3121 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3181 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3241 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     3301 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3361 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3421 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3481 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3541 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3601 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3661 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3721 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3781 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3841 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     3901 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     3961 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4021 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4081 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4141 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4201 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4261 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4321 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4381 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4441 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4501 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4561 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4621 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4681 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4741 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4801 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4861 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4921 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4981 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5041 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5101 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5161 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     5221 ttcatagccc atatatggag ttccg
//
