LOCUS       Exported                8220 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 8220)
  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..8220
                     /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-F
                     /note="Chimeric intron in CAG promoter, 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             4376..5137
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYMSGGLQKKLEELELD"
     primer_bind     complement(4421..4442)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4421..4437)
                     /label=pEGFP_N
     primer_bind     complement(4682..4701)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5029..5050
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5065..5081)
                     /label=EGFP CR
     primer_bind     5086..5138
                     /label=CNES Fwd
     primer_bind     complement(5090..5142)
                     /label=CNES Rvs
     CDS             5102..5134
                     /codon_start=1
                     /label=NES
                     /translation="LQKKLEELELD"
     primer_bind     complement(5104..5124)
                     /label=NES Colony R
     primer_bind     complement(5212..5231)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      5277..5332
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(5331..5350)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5723..5739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5723..5739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5736..5758)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5747..5763
                     /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(5771..5801)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5816..5837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5896..6091
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5942..6077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6004..6023
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(6027..6038)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    6097..6231
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6134..6153)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6188..6207
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6299..6316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6470..7058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6550..6569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7229..8089)
                     /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     7852..7871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8090..8194)
                     /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 attagcccgg
     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 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     4441 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     4501 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     4561 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     4621 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     4681 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     4741 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     4801 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     4861 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     4921 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     4981 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     5041 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca tgagcggcgg
     5101 cctgcagaag aagctggagg agctggagct ggactaatgt acaagtaaag cggccggccg
     5161 ccagcacagt ggtcgatcga ccaatgccct ggctcacaaa taccactgag atctttttcc
     5221 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     5281 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     5341 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     5401 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     5461 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     5521 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     5581 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     5641 tcttctctta tggagatccc tcgacctgca ccgtcgacca gctggtcgac ggtgcaccgt
     5701 cgaccagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5761 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5821 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5881 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5941 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6001 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6061 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     6121 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6181 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     6241 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6301 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6361 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6421 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6481 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6541 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6601 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6661 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6721 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6781 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6841 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6901 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6961 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7021 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7081 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7141 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7201 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7261 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7321 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7381 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7441 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7501 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7561 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7621 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7681 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7741 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7801 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7861 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7921 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7981 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8041 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8101 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8161 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
