LOCUS       pmRNA-iSyn-EGFP        7221 bp DNA     circular SYN 19-AUG-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7221)
  AUTHORS   Procurement Team
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7221
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     intron          647..1663
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
                     /note="color: #------"
     primer_bind     1720..1752
                     /label=circRNA F w/ T7 class III Phi-6.5 promoter
                     /note="color: black; sequence: 
                     AAAAAAAAAAAAAAAAAAAAAAAAAAAGGCCAGTGAATTGTAATACGACTCACTATAGG
                     G; added: 2024-07-03"
     promoter        1733..1752
                     /label=T7 class III Phi-6.5 promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        1733..1751
                     /label=T7 class III Phi-6.5 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #cc99ff; direction: RIGHT"
     intron          1809..2064
                     /label=T4 td upstream intron
                     /note="color: #------"
     exon            2041..3678
                     /label=circRNA exon
                     /note="color: #------"
     misc_feature    2056..2140
                     /label=mRNA 5′ UTR
                     /note="color: #ffff99"
     primer_bind     2056..2070
                     /label=mRNA F w/ A-inserted T7 class III Phi-6.5 promoter
                     /note="color: black; sequence: 
                     GACTTAATACGACTCACTATAAGgcgtcatattcggcg; added: 2024-07-03"
     misc_feature    2085..2134
                     /label=PABP motif
                     /note="color: #ccffff"
     misc_feature    2141..2806
                     /label=HRV-B3-derived synthetic IRES
                     /note="color: #ffcc99"
     CDS             join(2807..2809,2810..2812,2813..3526)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 2807 .. 2809 / #05fd14
                        2: 2810 .. 2812 / #05fd14 / 1a
                        3: 2813 .. 3526 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3527..3667
                     /label=mRNA 3′ UTR
                     /note="color: #ffff99"
     misc_feature    3536..3636
                     /label=HBA1 UTR
                     /note="color: #ccffff"
     primer_bind     complement(3652..3667)
                     /label=mRNA R w/ A120 tail
                     /note="color: black; sequence: 
                     TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
                     TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
                     TTcatctactgagtcggc; added: 2024-07-03"
     intron          3657..3863
                     /label=T4 td downstream intron
                     /note="color: #------"
     terminator      3923..3970
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     primer_bind     complement(3941..3970)
                     /label=circRNA R
                     /note="color: black; sequence: 
                     TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCAAAAAACCCCTCAAGACCCGTTTAGAGG
                     C; added: 2024-07-03"
     polyA_signal    3984..4208
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     protein_bind    4251..4267
                     /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)."
                     /note="color: #31849b"
     polyA_signal    4340..4461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     CDS             complement(4621..5220)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(5231..5559)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      5245..5380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             complement(5593..6408)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      6633..7221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      421 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      481 gggggggggg ggcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
      541 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
      601 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
      661 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
      721 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      781 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
      841 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
      901 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
      961 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
     1021 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
     1081 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
     1141 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
     1201 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
     1261 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
     1321 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
     1381 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1441 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     1501 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     1561 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1621 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1681 ggttattgtg ctgtctcatc attttggcaa agcggccgcg gccagtgaat tgtaatacga
     1741 ctcactatag gggttaacaa aaaaaaaaaa aaaaaaaaaa aatcaccgac cgatctatct
     1801 gaaccggtgg gaattctaga gaaaatttcg tctggattag ttacttatcg tgtaaaatct
     1861 gataaatgga attggttcta cataaatgcc taacgactat ccctttgggg agtagggtca
     1921 agtgactcga aacgatagac aacttgcttt aacaagttgg agatatagtc tgctctgcat
     1981 ggtgacatgc agctggatat aattccgggg taagattaac gaccttatct gaacataacg
     2041 ctaccgttta atattgcgtc atattcggcg accatttgtg tggtaaaaaa aaaaaaccaa
     2101 aaaaaaaaaa caaaaaaaaa aaataattga ctaagatatc ttaaaacagc ggatgggtac
     2161 cccaccatcc gacccactgg gtgtagtact ctggtacttc gtacctttgt acgcctgttc
     2221 ttcccattgt acccttcctg aacttccaac ccaagtaacg ttagaagctc aacatttagt
     2281 acaacaggaa gcaccacatc cagtggtgtt tagtacaagc acttctgttt ccccggagcg
     2341 aggtataggc tgtacccact gccaaaaacc tttaaccgtt atccgccaac caactacgta
     2401 aaagctagta gtattatgtt tttaactagg cgttcgatca ggtggatttc ccctccacta
     2461 gtttggtcga tgaggctagg aattccccac gggtgaccgt gtcctagcct gcgtggcggc
     2521 caacccagcc cactcactat ttgttttcgc gcccagttgc aaaaagtgtc ggggctggga
     2581 cgccttttta tagacatggt gtgaagactc gcatgtgctt ggttgtgatt cctccggccc
     2641 ctgaatgcgg ctaaccttaa ccctggagcc ttgtgtcaca aaccagtgat gataaggtcg
     2701 taatgagcaa ttccgggacg ggaccgacta ctttgggtgt ccgtgtttct tatttttctt
     2761 attattgtct tatggtcaca gcatatatat aacatatact gtgatcatgg tgagcaaggg
     2821 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2881 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2941 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3001 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     3061 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3121 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3181 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3241 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     3301 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3361 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     3421 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3481 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaataac aattggctgg
     3541 agcctcggtg gccatgcttc ttgccccttg ggcctccccc cagcccctcc tccccttcct
     3601 gcacccgtac ccccgtggtc tttgaataaa gtctgaacca cacaaatggt cgccgactca
     3661 gtagatgttt tcttgggtta attgaggcct gagtataagg tgacttatac ttgtaatcta
     3721 tctaaacggg gaacctctct agtagacaat cccgtgctaa attgtaggac tgccctttaa
     3781 taaatacttc tatatttaaa gaggtattta tgaaaagcgg aatttatcag attaaaaata
     3841 ctttctctag agtcgacctg cagacgcgtt cagatagatc ggtcggtgaa aaaaaaaaaa
     3901 aaaaaaaaaa aaaagcggcc gcctagcata accccttggg gcctctaaac gggtcttgag
     3961 gggttttttg gctagcggcc ggcctgtgcc ttctagttgc cagccatctg ttgtttgccc
     4021 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4081 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4141 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4201 ctctatggcg gcgcgccact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     4261 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     4321 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4381 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4441 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     4501 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     4561 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
     4621 tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc
     4681 ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtttc
     4741 caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct
     4801 gcccagaccc ttgccctggt ggtcgggcga cacgccgacg gtggccagga accacgcggg
     4861 ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga
     4921 accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct
     4981 ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat
     5041 gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg
     5101 gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg
     5161 tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat
     5221 gcgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     5281 gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     5341 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     5401 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     5461 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     5521 ctggggagcc tggggacttt ccacacccta actgacacag agctcgtgtt acaaccaatt
     5581 aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     5641 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     5701 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     5761 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     5821 atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg
     5881 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     5941 cattcgtgat tgcgcctgag cgaggcgaaa tacgcgatcg ctgttaaaag gacaattaca
     6001 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     6061 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     6121 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     6181 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     6241 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     6301 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     6361 atttaatcgc ggcctggagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     6421 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     6481 aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc
     6541 tgagttgaag gatcagtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6601 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6661 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6721 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6781 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6841 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6901 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6961 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7021 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7081 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7141 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7201 ggggggcgga gcctatggaa a
//
