LOCUS       Exported                4895 bp ds-DNA     circular SYN 25-JUN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCI_FLAG-GFP(39TAG)-6His
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4895)
  AUTHORS   Gemma
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 1, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     ORIGDB|GenBank
            Freezer Position: -20*C Plasmid Box #5: 28-30
FEATURES             Location/Qualifiers
     source          1..4895
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        295..674
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        675..878
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          1014..1146
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1191..1209
                     /gene="T7 prom"
                     /label=T7\prom
                     /note="Unknown feature type:Promoter"
                     /note="/vntifkey=21"
     regulatory      1213..1222
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1219..1980
                     /label=FLAG-GFP-6HIS
                     /note="/vntifkey=21"
     CDS             1222..1245
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1246..1959
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDAT*GKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             1960..1977
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2004..2125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2306..2761
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3093..3197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3198..4058
                     /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"
     rep_origin      4229..4817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caaavcttct catagacata catgtccaat tccatcaaav ctcctaactg aataactgcc
       61 ttvcggttaa tttttactta tttcggvata atgaaggtaa ctaaacggtc vtatacatta
      121 ttaacatact aacacccatt aacttattct ttaacattca atattggcca ttagccatat
      181 tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
      241 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca tgttggcatt
      301 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      361 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      421 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      481 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      541 atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      601 atgcccagta catgacctta cgggactttc ctacttggca gtacatctac gtattagtca
      661 tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg
      721 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      781 aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg
      841 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatca
      901 ctagaagctt tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg
      961 acacaacagt ctcgaactta agctgcagaa gttggtcgtg aggcactggg caggtaagta
     1021 tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa
     1081 gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt gcctttctct
     1141 ccacaggtgt ccactcccag ttcaattaca gctcttaagg ctagagtact taatacgact
     1201 cactataggc tagccaccat ggattacaaa gatgatgatg ataaagtgag caagggcgag
     1261 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1321 aagttcagcg tgtccggcga gggcgagggc gatgccacct agggcaagct gaccctgaag
     1381 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1441 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     1501 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1561 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1621 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1681 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggccaacttc
     1741 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1801 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1861 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1921 gccgccggga tcactctcgg catggacgag ctgtacaagc atcaccatca ccatcactga
     1981 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2041 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2101 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2161 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     2221 cgataaggat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2281 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2341 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2401 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2461 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2521 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2581 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2641 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2701 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     2761 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     2821 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     2881 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2941 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     3001 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3061 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3121 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3181 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3241 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3301 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3361 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3421 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3481 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3541 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3601 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3661 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3721 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3781 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3841 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3901 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3961 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4021 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4081 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4141 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4201 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4261 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4321 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4381 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4441 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4501 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4561 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4621 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4681 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4741 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4801 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4861 ggccttttgc tcacatggct cgacagatct vctca
//
