LOCUS       Exported                7417 bp ds-DNA     circular SYN 04-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7417)
  AUTHORS   Lena
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 28, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7417
                     /organism="recombinant plasmid"
                     /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     1729..1755
                     /label=JunoF-XhoI
     primer_bind     1735..1755
                     /label=Juno F NheI
     primer_bind     1735..1755
                     /label=JunoF-EcoRV
     CDS             1782..2516
                     /codon_start=1
                     /label=Juno
                     /translation="MAQWWQILLGLWAVLPTLAGDKLLSVCMNSKRHKQEPGPEDELYQ
                     ECRPWEDNACCTRSTSWEAHLEEPLLFNFSMMHCGLLTPACRKHFIQAICFHECSPNLG
                     PWIQPVVPNGQEEQRVWGVPLCQEDCEDWWRACHSSLTCKSNWLHGWDWSEEKKHCPAH
                     EPCLPFSYHFPTPDDLCEKIWNNTFKASPERRNSGRCLQKWFEPTLSNPNVEVALHFAG
                     SALAPQLSYTLPAFSLCLLFHP"
     primer_bind     complement(2581..2606)
                     /label=JunoR-NheI
     primer_bind     complement(2581..2602)
                     /label=Juno R-NotI
     primer_bind     complement(2581..2600)
                     /label=Juno R SmaI
     misc_feature    2652..3238
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2819..2836)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3046..3065
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3245..3622
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      3635..3644
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3641..4318
                     /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(3771..3789)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4039..4058
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4233..4256
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4409..4428)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      4474..4529
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(4528..4547)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4920..4936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4920..4936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4933..4955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4944..4960
                     /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(4968..4998)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5013..5034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5093..5288
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5139..5274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5201..5220
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5224..5235)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5294..5428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5331..5350)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5385..5404
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5496..5513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5667..6255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5747..5766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6426..7286)
                     /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     7049..7068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7287..7391)
                     /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 cgagctcttt
     1741 ggcatcagga ggagcaacag gagggccctg ccttgaaggt catggcacag tggtggcaga
     1801 tcctcttggg gttgtgggca gtcctaccca ccttggcagg ggacaaactg ctcagcgtct
     1861 gcatgaattc caagcgccac aagcaagaac ctggcccaga agacgaactc taccaggagt
     1921 gcaggccttg ggaggacaat gcctgctgca cacgttccac aagttgggaa gcccaccttg
     1981 aggagccctt gctctttaac ttcagcatga tgcactgtgg actgctgacc ccggcctgtc
     2041 gcaagcactt catccaggcc atctgcttcc atgagtgttc ccccaacctg gggccttgga
     2101 tccagccggt ggtcccaaac gggcaggaag agcagcgggt ttggggcgtg ccgctgtgcc
     2161 aggaggattg tgaggactgg tggcgagcct gccactcatc tttgacttgt aaatccaact
     2221 ggctccatgg ctgggactgg agtgaggaga agaaacactg ccccgcacat gagccctgcc
     2281 tccctttttc gtatcacttc cccaccccgg atgacctgtg tgagaagatc tggaacaaca
     2341 cattcaaggc cagtcccgag cgcaggaaca gtgggcggtg tctgcagaag tggtttgagc
     2401 ccacccttag caaccccaac gtggaagtgg ccctccactt tgccggctct gccttagccc
     2461 cacagctctc ttacaccctc ccagccttct ctctgtgcct gttgttccat ccctgatccc
     2521 tgcttccctc tccaacccac atttcctgat ccttctgggc catggccatc tccatcctca
     2581 ggctattcat gttgggggca gctagctcat cgataagctt gatatcgaat tcctgcagcc
     2641 cgggggatcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     2701 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     2761 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     2821 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     2881 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     2941 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     3001 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     3061 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     3121 gggcctcggt acacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     3181 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     3241 ggttatgcca gagccagcga agtctgctcc cgccccgaaa aagggctcca agaaggcggt
     3301 gactaaggcg cagaagaaag gcggcaagaa gcgcaagcgc agccgcaagg agagctattc
     3361 catctatgtg tacaaggttc tgaagcaggt ccaccctgac accggcattt cgtccaaggc
     3421 catgggcatc atgaattcgt ttgtgaacga cattttcgag cgcatcgcag gtgaggcttc
     3481 ccgcctggcg cattacaaca agcgctcgac catcacctcc agggagatcc agacggccgt
     3541 gcgcctgctg ctgcctgggg agttggccaa gcacgccgtg tccgagggta ctaaggccat
     3601 caccaagtac accagcgcta aggatccacc ggtcgccacc atggcctcct ccgaggacgt
     3661 catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg gccacgagtt
     3721 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     3781 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc agttccagta
     3841 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     3901 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     3961 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     4021 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac
     4081 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagatga ggctgaagct
     4141 gaaggacggc ggccactacg acgccgaggt caagaccacc tacatggcca agaagcccgt
     4201 gcagctgccc ggcgcctaca agaccgacat caagctggac atcacctccc acaacgagga
     4261 ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactccaccg gcgcctaagc
     4321 ggccgcaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     4381 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     4441 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     4501 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     4561 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     4621 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     4681 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     4741 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     4801 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     4861 acctgcaccg tcgaccagct ggtcgacggt gcaccgtcga ccagcttggc gtaatcatgg
     4921 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4981 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5041 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     5101 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5161 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5221 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5281 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5341 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5401 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     5461 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5521 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5581 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5641 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5701 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5761 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5821 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     5881 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5941 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6001 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6061 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6121 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6181 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6241 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6301 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6361 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6421 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6481 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6541 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6601 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6661 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6721 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6781 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6841 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6901 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6961 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7021 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7081 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7141 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7201 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7261 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7321 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7381 ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
