LOCUS       Exported                7926 bp ds-DNA     circular SYN 23-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7926)
  AUTHORS   Nico
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 28, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7926
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1783..1809
                     /label=mIzumo1 F NheI
     misc_feature    1832..3023
                     /label=Izumo1
     CDS             1841..3145
                     /codon_start=1
                     /label=Izumo1 tagged
                     /translation="MGPHFTLLLAALANCLCPGRPCIKCDQFVTDALKTFENTYLNDHL
                     PHDIHKNVMRMVNHEVSSFGVVTSAEDSYLGAVDENTLEQATWSFLKDLKRITDSDLKG
                     ELFIKELLWMLRHQKDIFNNLARQFQKEVLCPNKCGVMSQTLIWCLKCEKQLHICRKSL
                     DCGERHIEVHRSEDLVLDCLLSWHRASKGLTDYSFYRVWENSSETLIAKGKEPYLTKSM
                     VGPEDAGNYRCVLDTINQGHATVIRYDVTVLPPKHSEENQPPNIITQEEHETPVHVTPQ
                     TPPGQEPESELYPELHPELYPELIPTVAQNPEKKMKTRLLILLTLGFVVLVASIIISVL
                     HFRKVSAKLKNASDEVKPTASGSKSDQSLSQQMGLKKASQADFNSDYSGDKSEATENTR
                     TRPSSLEGPRFEGKPIPNPLLGLDSTRTGHHHHHH"
     primer_bind     complement(3035..3051)
                     /label=mIzumo1 Rvs
     CDS             3074..3115
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3116..3146)
                     /label=6HisStopSacI_anti
     primer_bind     complement(3116..3145)
                     /label=6HisStopXho_anti
     primer_bind     complement(3125..3156)
                     /label=Izumo R SmaI
     primer_bind     complement(3125..3147)
                     /label=6HisStopNheI_anti
     CDS             3125..3142
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3161..3747
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3328..3345)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3555..3574
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3754..4131
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      4144..4153
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4150..4827
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(4280..4298)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4548..4567
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4742..4765
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4918..4937)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      4983..5038
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(5037..5056)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5429..5445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5429..5445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5442..5464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5453..5469
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5477..5507)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5522..5543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5602..5797
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5648..5783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5710..5729
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5733..5744)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5803..5937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5840..5859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5894..5913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6005..6022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6176..6764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6256..6275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6935..7795)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7558..7577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7796..7900)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgct cgagagttcg
     1741 aagccttaag tacgcgtagg cgcgccttaa ttaacgttta aacgctagcg aattcggtcg
     1801 actggatccg gtaccgagga gatctgccgc cgcgatcgcc atggggccgc attttacact
     1861 cttgctggca gctcttgcca actgcctgtg tccagggagg ccctgcatca aatgtgacca
     1921 gtttgtgaca gatgcgctaa agactttcga aaacacttac ctgaatgacc acctgccaca
     1981 cgacattcac aaaaatgtaa tgaggatggt gaaccatgaa gtatcgagct tcggcgtagt
     2041 cacttcggct gaggattcct atttgggggc cgtggacgag aacacactgg aacaagcaac
     2101 ctggagtttt ctgaaggatc tgaagcgtat tacagacagt gacttaaaag gagagctctt
     2161 tataaaggaa ctattgtgga tgcttcgtca tcaaaaggac atctttaaca atcttgctag
     2221 acagttccaa aaggaagttc tttgtcccaa caaatgcgga gtgatgtcgc agactttgat
     2281 ctggtgtctt aagtgcgaaa agcagttgca catttgtcgg aaatccctag attgtggaga
     2341 gcgccacata gaggtacatc gctcggaaga cctggtcctg gactgtctgc tcagttggca
     2401 tcgtgcttct aagggactta cagattacag tttttacagg gtttgggaga acagttctga
     2461 gaccttgatt gccaagggga aagaaccata tctgaccaag tcgatggtgg gtccagagga
     2521 tgctggcaac taccgctgtg tgctagatac catcaaccaa ggtcatgcca ccgtcatccg
     2581 ctacgatgtc acagtattgc ccccaaagca ttcagaggaa aaccaaccac cgaacatcat
     2641 aacccaagag gagcacgaga ctcctgtcca cgtgactcca cagacaccac cggggcagga
     2701 gccagagtcg gagctgtacc cggagctgca cccagagttg tacccggagc tcatacctac
     2761 agtcgctcaa aatccagaga aaaaaatgaa aactcgcctc ttaatactgt tgacccttgg
     2821 ctttgtggtt cttgtggcca gcatcattat ctcggtactt cactttagga aagtcagcgc
     2881 taaattgaag aatgcaagtg acgaagtcaa acccaccgcg tcaggatcca agtcagatca
     2941 gagtttgtcc caacagatgg gattgaagaa ggcctctcag gcagatttta actctgacta
     3001 ctctggagat aagagcgagg caacagaaaa cacgcgtacg cggccctcga gtctagaggg
     3061 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     3121 cggtcatcat caccatcacc attgagcccc gggggatccg cccctctccc tccccccccc
     3181 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     3241 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     3301 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     3361 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     3421 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     3481 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     3541 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     3601 aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt
     3661 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     3721 aaacacgatg ataatatggc cacaaccatg gttatgccag agccagcgaa gtctgctccc
     3781 gccccgaaaa agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag
     3841 cgcaagcgca gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc
     3901 caccctgaca ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac
     3961 attttcgagc gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc
     4021 atcacctcca gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag
     4081 cacgccgtgt ccgagggtac taaggccatc accaagtaca ccagcgctaa ggatccaccg
     4141 gtcgccacca tggcctcctc cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc
     4201 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     4261 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
     4321 tgggacatcc tgtcccctca gttccagtac ggctccaagg cctacgtgaa gcaccccgcc
     4381 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     4441 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     4501 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     4561 aagaagacca tgggctggga ggcctccacc gagcggatgt accccgagga cggcgccctg
     4621 aagggcgaga tcaagatgag gctgaagctg aaggacggcg gccactacga cgccgaggtc
     4681 aagaccacct acatggccaa gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc
     4741 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc
     4801 gagggccgcc actccaccgg cgcctaagcg gccgcaattc actcctcagg tgcaggctgc
     4861 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     4921 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     4981 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     5041 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     5101 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
     5161 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     5221 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     5281 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     5341 tgtccctctt ctcttatgga gatccctcga cctgcaccgt cgaccagctg gtcgacggtg
     5401 caccgtcgac cagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5461 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5521 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5581 acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5641 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5701 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5761 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct
     5821 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5881 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc
     5941 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6001 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6061 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6121 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6181 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6241 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6301 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6361 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6421 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6481 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6541 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6601 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6661 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6721 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6781 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6841 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6901 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6961 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7021 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7081 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7141 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7201 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7261 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7321 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7381 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7441 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7501 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7561 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7621 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7681 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7741 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7801 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7861 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7921 cacctg
//
