LOCUS       Exported                8278 bp ds-DNA     circular SYN 25-APR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8278)
  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..8278
                     /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-F
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1782..1807
                     /label=EFF-1 Fwd NheI
     primer_bind     1790..1808
                     /label=EFF-1 F - EcoRV
     primer_bind     1790..1807
                     /label=EFF-1 for
     primer_bind     1790..1807
                     /label=Fwd Lena SpeI
     CDS             1791..3834
                     /codon_start=1
                     /label=EFF-1_V5
                     /translation="MEPPFEWSPQFILLLLAVTTYGFPLEEKFDGLFRAEPPHCSKTPI
                     VRAQTSQNAMSSIARGMQMQFSIGLHTAVCFRLYEDTQLASQEINDDENAGNQTSLLHT
                     IRLEKLEHHHPITQRYTFGIPEVHASCICECDATSSTCTAESHQFTACPESDKSDETSS
                     CYRTFFPNQTPIGCSEDDIPKLCCDVRFKPYKNMTFLAVKLEQPTTYATFVYAAYDFVN
                     GYWVEKDKTKIRSQLDGGTQDRHLDQKRRISLAVTAGGRASHQLETGMYFSRTSNGGET
                     EELRMQPLNEITDNNFDRLGWYRMDDSGHFHVNNGVVKMDDIHKAKVKNCKEQTYKSIL
                     SANHYMPGHFNLTRPLEVIKPWIQSARIFDSSLRQAVVTHAEGTNLQISIHLDDEVESQ
                     NLVFFHNASRIRDFSGSIIVDSKSNRLFNLTVYEASGKIDGSVKMSTGFGSDTIHTFTA
                     YVSDLHASNRSMIIPLPAIVGQGARAICLRADSMADIDKICHVIEYFESPLEIDLVEGK
                     WHEMIGTCPTCNQINFNGMMKFLNPAHWIKGISSIGDGVMIATDIVVYLGVLCILYLLI
                     TKIIVPLVRCWVCPMSIFCNGSSSSSKNKNDKRRKEREERRRKDKFVSESEDGARSSSE
                     PHDTLARYHGNHSERHYSSSQYIGLEGPRFEGKPIPNPLLGLDST"
     primer_bind     2547..2568
                     /label=EFF1 seq f
     CDS             3789..3830
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3814..3839)
                     /label=Reverse Liliana XmaI
     primer_bind     complement(3819..3839)
                     /label=EFF-1 R-NheI
     misc_feature    3847..4433
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4014..4031)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4241..4260
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4434..5195
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYMSGGLQKKLEELELD"
     primer_bind     complement(4479..4500)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4479..4495)
                     /label=pEGFP_N
     primer_bind     complement(4740..4759)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5087..5108
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5123..5139)
                     /label=EGFP CR
     primer_bind     5144..5196
                     /label=CNES Fwd
     primer_bind     complement(5148..5200)
                     /label=CNES Rvs
     CDS             5160..5192
                     /codon_start=1
                     /label=NES
                     /translation="LQKKLEELELD"
     primer_bind     complement(5162..5182)
                     /label=NES Colony R
     primer_bind     complement(5270..5289)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      5335..5390
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(5389..5408)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5781..5797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5781..5797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5794..5816)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5805..5821
                     /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(5829..5859)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5874..5895
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5954..6149
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6000..6135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6062..6081
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(6085..6096)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    6155..6289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6192..6211)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6246..6265
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6357..6374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6528..7116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6608..6627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7287..8147)
                     /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     7910..7929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8148..8252)
                     /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 aacgctagct atggaaccgc
     1801 cgtttgagtg gtctccccag tttattctgc ttctcctagc agttacaacc tacggatttc
     1861 cactcgagga gaaattcgat gggctcttcc gtgcagagcc accacactgt tccaagacac
     1921 caattgtccg tgctcaaaca tctcaaaatg cgatgtcatc aattgcaaga ggaatgcaaa
     1981 tgcaattctc aattggtttg cacactgcag tctgcttcag attatatgaa gacactcaac
     2041 tcgcatccca agagatcaat gatgatgaaa atgctggaaa tcagacatca cttcttcata
     2101 ctattcgatt ggaaaagctt gaacatcacc atccgataac tcagcggtac acttttggaa
     2161 tccctgaagt tcatgccagt tgtatctgtg agtgtgatgc cacatcttca acatgtaccg
     2221 cagaatctca tcagtttacc gcttgtccag agtctgataa gtctgatgaa acatcttcat
     2281 gctaccgcac gttcttccca aatcagacac caatcgggtg cagtgaagac gacatcccca
     2341 agctttgctg tgacgttcga ttcaagccat ataaaaatat gacattcctc gcagtcaaac
     2401 tagaacaacc tacaacgtat gcaacatttg tctacgcagc ttatgatttt gtaaatggat
     2461 attgggtgga aaaagataaa actaaaatcc gttctcaatt agatggtgga acacaggata
     2521 ggcatctcga ccaaaaacgt cgaatttcat tggctgtaac tgccggagga cgagcatctc
     2581 atcaactgga gacaggaatg tatttctcaa gaacaagtaa tggaggagaa acagaagaac
     2641 tgagaatgca accattgaat gagattacag acaataactt tgaccgtctt ggatggtaca
     2701 gaatggacga ttctggtcat tttcatgtga ataatggagt tgtgaaaatg gatgatattc
     2761 acaaggctaa agtgaaaaat tgcaaagagc aaacttataa atcaattctc tctgcaaatc
     2821 attatatgcc aggacatttc aatttgactc gtccacttga agttatcaaa ccgtggattc
     2881 aatctgctcg aatttttgat agttctctca gacaagctgt tgtaacacat gccgaaggaa
     2941 ctaatcttca aatttcaatt cacttggatg acgaggtgga aagtcaaaac cttgtcttct
     3001 tccataatgc atcaagaatt cgtgacttca gtggatccat catcgttgat tcaaaatcaa
     3061 atcgattgtt caacttgaca gtatatgagg cttctggaaa aattgatgga tcagtgaaga
     3121 tgtcaactgg atttggatct gatacaattc acacattcac tgcttatgtt agtgatcttc
     3181 atgcttcaaa ccgttctatg attattccac tgccagccat cgttggacaa ggagccagag
     3241 caatctgtct ccgagcagat tcaatggctg atattgacaa aatttgtcat gtcattgagt
     3301 atttcgagag ccctttggaa atcgatcttg ttgagggaaa atggcacgaa atgattggaa
     3361 cctgcccaac ttgtaatcaa atcaatttca atggaatgat gaaatttctg aatccagctc
     3421 attggatcaa aggaatcagt tcgattggtg acggcgttat gattgctacg gatattgttg
     3481 tgtatctcgg agttttgtgt attctgtatc ttcttattac gaagattatt gttccgttgg
     3541 ttcgttgctg ggtatgcccc atgtcgattt tctgcaatgg gtcctcttcg tcgtccaaaa
     3601 acaaaaatga taagagaagg aaggagagag aggagaggcg cagaaaggac aaatttgtct
     3661 cagaatccga agatggtgca agaagtagtt ccgagcccca cgacactttg gcgagatacc
     3721 atggaaatca ctcggagcgg cactatagca gtagccagta cattggtcta gagggcccgc
     3781 ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg tgacccgggg
     3841 gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     3901 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     3961 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     4021 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     4081 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     4141 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     4201 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     4261 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     4321 tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     4381 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga
     4441 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4501 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     4561 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     4621 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     4681 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     4741 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     4801 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     4861 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     4921 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     4981 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     5041 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     5101 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacatg agcggcggcc
     5161 tgcagaagaa gctggaggag ctggagctgg actaatgtac aagtaaagcg gccggccgcc
     5221 agcacagtgg tcgatcgacc aatgccctgg ctcacaaata ccactgagat ctttttccct
     5281 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     5341 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     5401 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     5461 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     5521 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     5581 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     5641 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     5701 ttctcttatg gagatccctc gacctgcacc gtcgaccagc tggtcgacgg tgcaccgtcg
     5761 accagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5821 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5881 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5941 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6001 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6061 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6121 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     6181 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6241 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     6301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6361 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6421 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6721 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6961 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7201 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7861 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7921 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7981 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8041 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8101 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8221 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
