LOCUS       Exported                4911 bp ds-DNA     circular SYN 17-OCT-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Oct 18, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     SnapGene NotesSequence Type: SyntheticConfirmed Experimentally: 
            0Transformed Into: unspecifiedSnapGene UUID: 
            84f62078-7d26-4ae6-968d-bae7e8235c4fSequence Document Type: 
            UNAImported FromPath: 
            G:\.shortcut-targets-by-id\1MocL5GBiP9pjqqc00kp67EYa6ACf2MZf\Hsu 
            Lab\Plasmid mapsFilename: pCB233 - BB for pseudosite assay - 
            modified for NGS.dna
FEATURES             Location/Qualifiers
     source          1..4911
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        32..136
                     /gene="<i>bla</i>"
                     /label=AmpR promoter
     CDS             137..997
                     /codon_start=1
                     /transl_table=0
                     /gene="<i>bla</i>"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="/Original 
                     Translation=MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIEL
                     DLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPV
                     TEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEP
                     ELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALP
                     AGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLI
                     KHW*"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1168..1756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="/snapgene_direction=RIGHT"
     misc_feature    1969..2072
                     /label=Cp36 attD
                     /note="/created by=aliso"
     unsure          2020..2021
                     /label=core
                     /note="/created by=aliso"
     gap             2077..2088
                     /estimated_length=12
     promoter        2117..3294
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2348..3285
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     misc_feature    3296..3305
                     /label=Kozak seq
     CDS             3302..3898
                     /codon_start=1
                     /transl_table=0
                     /gene="<html><body><i>pac</i> from <i>Streptomyces 
                     alboniger</i></body></html>"
                     /product="<html><body>puromycin 
                     <i>N</i>-acetyltransferase</body></html>"
                     /label=PuroR
                     /note="<html><body>confers resistance to 
                     puromycin</body></html>"
                     /note="/Original 
                     Translation=MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVT
                     ELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQ
                     QQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAP
                     RNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             3899..3955
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1  
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding 
                     separatepolypeptides."
                     /note="/Original Translation=ATNFSLLKQAGDVEENPGP"
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3962..4672
                     /codon_start=1
                     /transl_table=0
                     /gene="mCherry"
                     /product="red fluorescent protein mCherry"
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein"
                     /note="codon-optimized for expression in mammalian cells"
                     /note="/dnas_title=mCherry"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    4703..4910
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
       61 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      121 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      181 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      241 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      301 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      361 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      421 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      481 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      541 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      601 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      661 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      721 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      781 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      841 cggtgagcgt ggctctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      901 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      961 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1021 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1081 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1141 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1201 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1261 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1321 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1381 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1441 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1501 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1561 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1621 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1681 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1741 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1801 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1861 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1921 gagcgaggaa gcggaagagc gagcttcgca cgattgcacc ttgggagcct taaccgcttt
     1981 tgaatgtccg tcttagttaa tccctaattg aacgctccaa aaggtgactt ccaataggga
     2041 tttattcctt ttaaaattaa cggcataatc gtgccgnnnn nnnnnnnngc cgccgaatgc
     2101 aacctcgatc gagccgggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     2161 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
     2221 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     2281 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     2341 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
     2401 ttgcgtgcct tgaattactt ccactggctg cagtacgtga ttcttgatcc cgagcttcgg
     2461 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
     2521 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
     2581 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
     2641 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
     2701 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
     2761 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
     2821 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
     2881 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
     2941 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     3001 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     3061 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     3121 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     3181 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     3241 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaagccac
     3301 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     3361 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     3421 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     3481 catcggcaag gtgtgggtcg cggacgacgg cgccgccgtg gcggtctgga ccacgccgga
     3541 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     3601 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     3661 gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg
     3721 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     3781 ggaaacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     3841 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgc
     3901 cacgaacttc tctctgttaa agcaagcagg tgacgtggaa gaaaaccccg gtcccggatc
     3961 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     4021 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     4081 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     4141 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     4201 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     4261 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     4321 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     4381 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     4441 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     4501 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     4561 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     4621 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aactcgagct
     4681 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4741 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4801 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4861 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga c
//
