LOCUS       Exported                4773 bp ds-DNA     circular SYN 23-JUN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4773)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 25, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4773
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          402..459
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2440..2533
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     379..395
                     /label=M13 fwd
     misc_feature    402..418
                     /label=T7 promoter
     misc_feature    419..459
                     /label=HBA1-derived 5UTR
                     /note="CleanCap AG is optimized for production of mRNA that
                     initiates with AGG, conventional transcripts start with 
                     5'-GGG.
                     
                     
                     Best T7 promoter sequence would be 
                     5'TAATACGACTCACTATAAGGNNNNNNNNNNNN 3'  Using that sequence 
                     with the CleanCap AG reagent, N-7113, you will get 95-98% 
                     capped mRNA with the natural CAP1 structure
                     
                     
                     https://www.tebu-bio.com/blog/tips-to-modify-mrna-for-in-vi
                     vo-applications-and-delivery-optimisation/"
     misc_feature    460..559
                     /label=LHA
     5'UTR           560..734
     polyA_signal    741..789
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(953..1672)
                     /codon_start=1
                     /label=GFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(1745..1948)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        1949..2252
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     3'UTR           2259..2339
     misc_feature    2340..2439
                     /label=RHA
     misc_feature    2440..2533
                     /label=alpha-globin-3UTR
     primer_bind     complement(2552..2568)
                     /label=M13 rev
     protein_bind    2576..2592
                     /label=lac operator
     promoter        complement(2600..2630)
                     /label=lac promoter
     rep_origin      complement(2954..3542)
                     /direction=LEFT
                     /label=ori
     CDS             complement(3713..4573)
                     /codon_start=1
                     /label=AmpR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4574..4678)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ctaatacgac tcactataag
      421 gaaataagag agaaaagaag agtaagaaga aatataagag ttgacgcgat gtgatttctg
      481 cccagtgctc tgaatgtcaa agtgaagaaa ttcaatgaag cgcgggtaaa cggcgggagt
      541 aactatgact ctcttaaggg tctagttaca actgggcatc gctgcagaga tcgcacctcc
      601 tcgtggtccc gctggtagcc cttcgaaggg tgactaagtc gatctctgcc ccaggtacgg
      661 agccgttggg actcaccagt ccaacgtaac tcctgcctaa attcggtgaa acaaattcct
      721 cggtaaaaag ccccggtacc gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt
      781 ttattgccga tcccctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
      841 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
      901 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtcggccg ctttacttgt
      961 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     1021 gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg tagtggttgt
     1081 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggccagct
     1141 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     1201 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     1261 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     1321 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     1381 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     1441 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     1501 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     1561 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     1621 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     1681 attcgaagct tgagctcgag atctgagtcc ggtagcgcta gcggatctga cggttcacta
     1741 aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     1801 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     1861 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     1921 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     1981 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     2041 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     2101 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     2161 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     2221 caggcgggcc atttaccgta agttatgtaa cgcccgggca cgggatttgt ccaaggtgga
     2281 cgggccacct ttacttaacc cggaaaagga acatatataa tttatgtgtg ttcgataaat
     2341 agccaaatgc ctcgtcatct aattagtgac gcgcatgaat ggatgaacga gattcccact
     2401 gtccctacct actatccagc gaaaccacag ccaagggaaa gctgccttct gcggggcttg
     2461 ccttctggcc atgcccttct tctctccctt gcacctgtac ctcttggtct ttgaataaag
     2521 cctgagtagg aagaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2581 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2641 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2701 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2761 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2821 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2881 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2941 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3001 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3061 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3121 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3181 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3241 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3301 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3361 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3421 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3481 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3541 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3601 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3661 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3721 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3781 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3841 caatgatacc gcgtgagcca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3901 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3961 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4021 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4081 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4141 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4201 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4261 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4321 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4381 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4441 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4501 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4561 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4621 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4681 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4741 ataaaaatag gcgtatcacg aggccctttc gtc
//
