LOCUS       Exported                8554 bp ds-DNA     circular SYN 24-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8554)
  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..8554
                     /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"
     misc_feature    1790..3703
                     /label=Fsx1
     CDS             3704..3745
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3755..3772
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3789..4375
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3956..3973)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4183..4202
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4382..4759
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      4772..4781
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4778..5455
                     /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(4908..4926)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5176..5195
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5370..5393
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5546..5565)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      5611..5666
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(5665..5684)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6057..6073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6057..6073)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6070..6092)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6081..6097
                     /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(6105..6135)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6150..6171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6230..6425
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6276..6411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6338..6357
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(6361..6372)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    6431..6565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6468..6487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6522..6541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6633..6650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6804..7392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6884..6903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7563..8423)
                     /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     8186..8205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8424..8528)
                     /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 aacgctagca tgagacgtgc
     1801 agcattgatt ctggctttcg tactgttcat cggtctttcc tcggctactg ttacctcggc
     1861 tgattcaatc acgtataact ctggaaccag cgagttcttc gacggagatg tctttgcaat
     1921 agaggtaacc gctgaccagt cgactgatga gatagatatc tacctcggtg cttctgagct
     1981 ttcggaaaag acggacggcg aggtcaacca ggacctcagt attgagttca cgcaccagga
     2041 ttccaagctg aagtattcta cttcgacttc tgacgagcta cgggacattg tcactcttac
     2101 aacttactac gaggacgggt ttgacacgga gcaggatgct atagacgcca tcaaatcaga
     2161 ctgttacgat ctgaatcaga acgggaatgg aagcggaagg tacagcaggt actactcagt
     2221 tacatctcct gtctatgatt acgagatcta ttgtttccag aagaacgaga aactggctac
     2281 tccagcatac atagacaacc cggatgagat cttcactgca aaggctgagc tacaggccgg
     2341 tgacaagacg attcagtcag ctacattgtc taacggtgac gccggagacg gaacagtcac
     2401 tgacctcggt gactcaaaga ttagctggaa cgggaacctg gatttaggag cttctgaacc
     2461 ggagaatagc cgagtcattg ctttgtactc gaatgatttt gagaacgggt ggaggatcgg
     2521 caataagcag tcgtacgagg actacaagac gttcatcggt ggcggggatg cctacgatct
     2581 cctgattgac tggcaggacg ggacttacac cgcctctgaa gtagaggatg agctggtgaa
     2641 tactgatgcg aaccaggctg ttgaggaggc cagttctagc acgacagact tggtgaacgc
     2701 gaaggttaag gactcgtctc ttgacaccgg ttcgttcgtg tatgataccc ctgagctgct
     2761 ttcctacccg tccttcacag tctacgtcga tgccggagaa aacggataca tcgaggtaac
     2821 gaagccgact ggtgaccccg atatcatctc cacttcatct acagaaatta aggagggaga
     2881 tgagggaact gtaaccgcca ccgttgagaa tgtcggagat ggtgagggcg agttctctgg
     2941 aaggctttcc agctgcggtg aaggcttcag tattgtcgac gaccagaata cgaagaatgt
     3001 gggtgccgga gaatcggtga catacagttt tgatgtcgct ttcagttctg tcagcagtga
     3061 gtcgaaggag atctcgggca gctgcacgtt cgaggtgaac ggcgtcgagt ccagtgattc
     3121 caccagtgtt tctgtgaccg gtatccagca gagtgagtgt aatccaggcg atcagagacg
     3181 ggaaaagaat gagaatgacc gctgggagat ctacacctgc caggacaacg gcctgaccta
     3241 cgaatatgat gtaacttgtg cagaggacga gaaggctgta gctcagggag ataaccagtt
     3301 ttcctgtgag aaacaggacg atgattcagg tggcggagac aacactggat ctgactccgg
     3361 actgttctcg aatctgttcg gaggttctgg ttccggagat ctgcttacac aggttcacac
     3421 agctctgtct atcctcgcgg ggcttgttgc aggcttcttc ggttatagag gagcccgatg
     3481 gattcacggc gagactgata taaaaggagg cttcaagctg gagtcccgga acgtaagccg
     3541 tgttaagaga ggtagtcctg tagccggtat tgtcggagct gtactcgggt ttgtcgtagg
     3601 atacggtgtt gcctcagtct tccatccagt agttcagatt atagtggttc tgggaattgc
     3661 tgttggcttg tactatttca ggctagaggg cccgcggttc gaaggtaagc ctatccctaa
     3721 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgacccgg
     3781 gggatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     3841 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     3901 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     3961 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     4021 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     4081 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     4141 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     4201 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     4261 cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     4321 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt
     4381 tatgccagag ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga aggcggtgac
     4441 taaggcgcag aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga gctattccat
     4501 ctatgtgtac aaggttctga agcaggtcca ccctgacacc ggcatttcgt ccaaggccat
     4561 gggcatcatg aattcgtttg tgaacgacat tttcgagcgc atcgcaggtg aggcttcccg
     4621 cctggcgcat tacaacaagc gctcgaccat cacctccagg gagatccaga cggccgtgcg
     4681 cctgctgctg cctggggagt tggccaagca cgccgtgtcc gagggtacta aggccatcac
     4741 caagtacacc agcgctaagg atccaccggt cgccaccatg gcctcctccg aggacgtcat
     4801 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga
     4861 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     4921 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg
     4981 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     5041 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     5101 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     5161 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
     5221 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa
     5281 ggacggcggc cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca
     5341 gctgcccggc gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta
     5401 caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg cctaagcggc
     5461 cgcaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     5521 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     5581 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     5641 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     5701 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     5761 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     5821 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     5881 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     5941 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     6001 tgcaccgtcg accagctggt cgacggtgca ccgtcgacca gcttggcgta atcatggtca
     6061 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6121 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6181 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     6241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6361 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6421 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6541 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     6601 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6961 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     7021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7201 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8521 gttccgcgca catttccccg aaaagtgcca cctg
//
