LOCUS       Exported                8439 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 8439)
  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..8439
                     /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"
     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             4595..5356
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYMSGGLQKKLEELELD"
     primer_bind     complement(4640..4661)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4640..4656)
                     /label=pEGFP_N
     primer_bind     complement(4901..4920)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5248..5269
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5284..5300)
                     /label=EGFP CR
     primer_bind     5305..5357
                     /label=CNES Fwd
     primer_bind     complement(5309..5361)
                     /label=CNES Rvs
     CDS             5321..5353
                     /codon_start=1
                     /label=NES
                     /translation="LQKKLEELELD"
     primer_bind     complement(5323..5343)
                     /label=NES Colony R
     primer_bind     complement(5431..5450)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      5496..5551
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(5550..5569)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5942..5958)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5942..5958)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5955..5977)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5966..5982
                     /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(5990..6020)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6035..6056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6115..6310
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6161..6296
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6223..6242
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(6246..6257)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    6316..6450
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6353..6372)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6407..6426
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6518..6535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6689..7277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6769..6788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7448..8308)
                     /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     8071..8090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8309..8413)
                     /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 aaccatggtg agcaagggcg aggagctgtt
     4621 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4681 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     4741 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     4801 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4861 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     4921 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4981 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     5041 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     5101 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     5161 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     5221 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     5281 gatcactctc ggcatggacg agctgtacat gagcggcggc ctgcagaaga agctggagga
     5341 gctggagctg gactaatgta caagtaaagc ggccggccgc cagcacagtg gtcgatcgac
     5401 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     5461 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     5521 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     5581 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     5641 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     5701 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     5761 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     5821 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     5881 cgacctgcac cgtcgaccag ctggtcgacg gtgcaccgtc gaccagcttg gcgtaatcat
     5941 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6001 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6061 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct
     6121 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6181 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6241 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6301 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6361 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6421 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg
     6481 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     6901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8401 taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
