LOCUS       Exported                6869 bp ds-DNA     circular SYN 06-APR-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6869)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Apr 6, 2022 from SnapGene 4.3.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6869
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /label=source:recombinant plasmid
                     /note="/locus_tag=source:recombinant plasmid"
     source          1625..2359
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=color: #ffffff
                     /note="/locus_tag=color: #ffffff"
     enhancer        202..581
                     /locus_tag="CMV enhancer"
                     /label=CMV enhancer
                     /note="CMV enhancer"
     promoter        582..785
                     /locus_tag="CMV"
                     /label=CMV
                     /note="CMV"
     promoter        830..848
                     /locus_tag="T7"
                     /label=T7
                     /note="T7"
     misc_feature    862..896
                     /locus_tag="MCS"
                     /label=MCS
                     /note="MCS"
     misc_feature    902..1609
                     /locus_tag="mCherry"
                     /label=mCherry
     misc_feature    1610..1624
                     /locus_tag="(GGGGS)2 linker"
                     /label=GGGGS TAGlinker
                     /label=(GGGGS)2 linker
     misc_feature    1613..1615
                     /label=TAG
     CDS             1625..2359
                     /codon_start=1
                     /locus_tag="eGFP"
                     /label=eGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYKHHHHHH"
     primer_bind     1625..1647
                     /label=pCMV-eGFP-wt-F
     primer_bind     complement(2333..2359)
                     /label=pCMV-eGFP-wt-R
     CDS             2339..2356
                     /codon_start=1
                     /locus_tag="6xHis"
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2361..2410
                     /locus_tag="MCS(1)"
                     /label=MCS
                     /label=MCS(1)
                     /note="MCS"
     polyA_signal    2436..2660
                     /locus_tag="bGH"
                     /label=bGH
                     /note="bGH"
     rep_origin      2706..3134
                     /locus_tag="f1 ori"
                     /label=f1 ori
                     /note="f1 ori"
     promoter        3148..3477
                     /locus_tag="SV40"
                     /label=SV40
                     /note="SV40"
     rep_origin      3328..3463
                     /locus_tag="SV40 ori"
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             3544..4338
                     /codon_start=1
                     /locus_tag="G418"
                     /label=G418
                     /note="G418"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4512..4633
                     /locus_tag="SV40 poly(A) signal"
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
     rep_origin      complement(5084..5669)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="ori"
     CDS             complement(5840..6700)
                     /codon_start=1
                     /locus_tag="AmpR"
                     /label=AmpR
                     /note="AmpR"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6701..6805)
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
                     /note="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
      961 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1021 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     1081 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     1141 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     1201 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     1261 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     1321 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     1381 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     1441 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     1501 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     1561 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg gttagggtgg
     1621 tagcgtgagc aaaggcgaag aactgttcac cggtgtggtg ccgattctgg ttgaactgga
     1681 cggcgatgtg aacggccaca aattcagcgt gagtggcgaa ggtgaaggcg atgcgaccta
     1741 cggcaaactg acgctgaagt tcatctgcac cacgggcaaa ctgccggttc catggccgac
     1801 gctggtgacc acgctgacct atggcgtgca atgcttcagt cgctacccgg accacatgaa
     1861 gcagcacgac ttcttcaaaa gcgcgatgcc agaaggctac gtgcaagaac gcaccatctt
     1921 cttcaaggac gacggcaact acaaaacccg cgccgaggtg aaattcgagg gtgatacgct
     1981 ggtgaaccgc atcgagctca agggcatcga cttcaaagaa gacggcaaca ttctgggcca
     2041 caagctggag tataactaca acagccacaa cgtgtacatc atggcggaca agcagaaaaa
     2101 cggcatcaaa gttaacttca agatccgcca caacatcgag gatggcagcg tgcagctggc
     2161 cgatcattac cagcagaata cgccgattgg cgacggtccg gttctgctgc cagataacca
     2221 ctatctgagc acgcagagcg cgctgagcaa agatccgaac gagaagcgcg atcacatggt
     2281 tctgctggaa ttcgtgaccg ccgcgggcat caccctcggt atggatgaac tgtacaaaca
     2341 tcatcatcat catcattaag aattctgcag atatccagca cagtggcggc cgctcgagtc
     2401 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2461 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2521 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2581 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2641 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2701 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2761 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2821 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2881 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2941 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3001 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3061 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3121 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3241 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3301 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3361 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3421 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3481 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3541 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3601 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3661 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3721 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3781 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3841 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3901 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3961 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4021 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4081 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4141 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4201 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4261 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4321 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4381 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4441 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4501 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4561 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4621 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4681 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4741 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4801 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4861 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4921 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4981 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5041 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5101 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5161 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5221 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5281 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5341 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5401 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5461 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5521 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5581 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     5641 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5701 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5761 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5821 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5881 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5941 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6001 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6061 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6121 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6181 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6241 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6301 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6361 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6421 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6481 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6541 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6601 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6661 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6721 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6781 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     6841 gatcgggaga tctcccgatc ccctatggt
//
