LOCUS       Exported                8773 bp ds-DNA     circular SYN 04-APR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8773)
  AUTHORS   Tivan25
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 28, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8773
                     /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     2389..2408
                     /label=LXH-38
     primer_bind     3220..3239
                     /label=LXH-49
     CDS             3923..3964
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3974..3991
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    4008..4594
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4175..4192)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4402..4421
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4601..4978
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      4991..5000
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4997..5674
                     /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(5127..5145)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5395..5414
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5589..5612
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5765..5784)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      5830..5885
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(5884..5903)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6276..6292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6276..6292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6289..6311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6300..6316
                     /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(6324..6354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6369..6390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6449..6644
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6495..6630
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6557..6576
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(6580..6591)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    6650..6784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6687..6706)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6741..6760
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6852..6869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7023..7611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7103..7122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7782..8642)
                     /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     8405..8424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8643..8747)
                     /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 gtaccatggt
     1801 gaacgcgatt ttaatggctt gtatactcgc cggaattttc gtcggaatgt tcaacgaagt
     1861 cgacggaatt cagattttat cgaaatcaaa gctcgaaaag tgtgagaaaa catcagattc
     1921 tggtaatctc aattgttcca ctaagatcgt cttgaatctc gctgttccca gtggatccag
     1981 tggaggagag gcttcgattg tagcggagat cgtggaggtt gaagataact cgagtagtaa
     2041 tatgcaaacc gtacggattc caccggttat tacagtgaac aaatctgcag catatgctct
     2101 ctatgatctt acttacatta gggatgtacc ttataaacct caagagtatc atgtcacgac
     2161 tcgaaagtgt gaacccgatg ctggaccgga tattgtacaa atttgtgaga ggttgagaga
     2221 tgagaagggt aatgttttag agcagactca gccgatttgt tgtccatgtg gaccgcaacg
     2281 aagaatgcct tcttcttgtg gagatatttt tgataagatg attaaaggga aggcaaatac
     2341 cgcgcattgc cttcgttttc caggtgactg gttccatgtt ttcgggattg gacaacggtc
     2401 actaggattt agtgtgcgag ttgagttgaa aacaggaact agagtttcag aagtgatcat
     2461 tggtcctgaa aataggacag ctactgcaaa tgataacttt ctcaaagtta atctcatagg
     2521 agactttggt ggttacacaa gtatcccttc tttcgaggac ttctatcttg taatcccaag
     2581 agaggcggct gaagcgggac aaccagggag cttaggcgca aattactcaa tgtggatgct
     2641 gcttgagaga gtgagattta cattggatgg cttagaatgt aataaaatcg gtgttggtta
     2701 tgaagctttc aacacccaac caaatttctg ctcttctcca tattggagtt gcttgcataa
     2761 tcaactatgg aacttccgtg agtcagatat aaacagaatc gaccgacacc aacttccttt
     2821 atatggactt gaaggaagat ttgagaggat caatcaacac ccaaacgcgg ggcctcactc
     2881 gttctcaatt ggagtcacgg aaacactcaa cacaaatcta atgatagaac taagagctga
     2941 tgatattgaa tacgttttcc agaggagtcc cggtaagatc ataaacatcg cgataccgac
     3001 attcgaagca cttacacaat tcggagttgc tgcagtcata atcaagaata ctggtgaagt
     3061 agaagcgtct tatagcttaa cgtttgattg ctccaaaggc gtggcttttg ttgaggaaca
     3121 atttttcatt ataaaaccga aagcggttac tactcggtct tttaagctgt acccgacgaa
     3181 agatcaggct gcaaagtata tatgcacagc tatactcaag gactctcagt tcagcgaagt
     3241 agaccgagcc gagtgtcaat ttagtacaac ggctaccgta cttgacaatg gaactcaggt
     3301 tactaatcca tttcaaattc cagagactca acctaaaggt ttctttgatt cgatcaggat
     3361 actatggatg aaaatcatca atggtttagt cgatttcatc accggagata cttgcagaaa
     3421 caaatgctcg agtttcttcg acttcagttg ccacattcaa tacgtttgct taagctggat
     3481 ggttatgttt ggtcttctcc tcgctctctt cccaattact tgtttgcttc tttggctctt
     3541 gcaccaaaag ggactctttg atccatgcta cgactggtgg gaagatcatt tcgatttaga
     3601 ccatcatcgg agattattac cctccagagc agacgttgtt aaccgccatc atcaccatca
     3661 caagcatcgt catcaccata atcaccatcg taggacacac cagagacaca aacatcatca
     3721 tggtcaagac gacgacgtat tgcagaagat gatgcttgaa cgtgaccaca gcgactctca
     3781 ctactaccac caacttcatc gagtccataa agatagcaag cagaagcagc gtaggcgagc
     3841 caagcacggg atcgtcctgc cacgtgatgt gcacgtggaa agacagagga aacaaagact
     3901 acgtgagagt ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     3961 tacgcgtacc ggtcatcatc accatcacca ttagcccggg ggatccgccc ctctccctcc
     4021 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     4081 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     4141 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     4201 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     4261 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     4321 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     4381 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     4441 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     4501 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     4561 ctttgaaaaa cacgatgata atatggccac aaccatggtt atgccagagc cagcgaagtc
     4621 tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg
     4681 caagaagcgc aagcgcagcc gcaaggagag ctattccatc tatgtgtaca aggttctgaa
     4741 gcaggtccac cctgacaccg gcatttcgtc caaggccatg ggcatcatga attcgtttgt
     4801 gaacgacatt ttcgagcgca tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg
     4861 ctcgaccatc acctccaggg agatccagac ggccgtgcgc ctgctgctgc ctggggagtt
     4921 ggccaagcac gccgtgtccg agggtactaa ggccatcacc aagtacacca gcgctaagga
     4981 tccaccggtc gccaccatgg cctcctccga ggacgtcatc aaggagttca tgcgcttcaa
     5041 ggtgcgcatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     5101 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     5161 cttcgcctgg gacatcctgt cccctcagtt ccagtacggc tccaaggcct acgtgaagca
     5221 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     5281 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     5341 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     5401 aatgcagaag aagaccatgg gctgggaggc ctccaccgag cggatgtacc ccgaggacgg
     5461 cgccctgaag ggcgagatca agatgaggct gaagctgaag gacggcggcc actacgacgc
     5521 cgaggtcaag accacctaca tggccaagaa gcccgtgcag ctgcccggcg cctacaagac
     5581 cgacatcaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     5641 gcgcgccgag ggccgccact ccaccggcgc ctaagcggcc gcaattcact cctcaggtgc
     5701 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     5761 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     5821 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     5881 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     5941 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     6001 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     6061 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     6121 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     6181 tcatagctgt ccctcttctc ttatggagat ccctcgacct gcaccgtcga ccagctggtc
     6241 gacggtgcac cgtcgaccag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6301 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6361 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6421 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
     6481 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6541 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     6601 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
     6661 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6721 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6781 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6841 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6901 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6961 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7021 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7081 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7141 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7201 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7261 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7321 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7381 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7441 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     7501 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7561 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7621 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7681 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7741 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7801 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7861 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7921 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7981 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8041 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8101 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8161 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8221 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8281 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8341 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8401 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8461 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8521 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8581 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8641 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8701 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8761 aaagtgccac ctg
//
