LOCUS       Exported                7121 bp ds-DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7121)
  AUTHORS   Huang Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Apr 18, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7121
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             746..1606
                     /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"
     terminator      1619..1755
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1907..1978
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1983..2029)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2047..2095
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2096..2118)
                     /label=KHDG106 R
     primer_bind     2119..2143
                     /label=KHDG105 F
     enhancer        2176..2479
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2481..2683
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2767..3198
                     /codon_start=1
                     /label=Rhobin2
                     /translation="EKNQFIIDIYEKLNEYHKKLLELARALAEALRAGDRARAGRLLEE
                     FLATFKAMKAYGDAKEVESFLYFSEEELAAIFAESDVERKKLGVTLKIPEVVAMVEELG
                     RAFEAGDTATAAAIVEKLVKFYEANIIIGQKRIDELKSKI"
     CDS             3229..3516
                     /codon_start=1
                     /label=SEC61B
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     misc_feature    3527..4101
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3551..4103
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4121..4141
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4166..4864
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             4874..4921
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4931..4951
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5004..5125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      5240..5286
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5316..5452
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5497..5568
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(6533..7121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
      241 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
      301 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
      361 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
      421 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
      481 tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
      541 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
      601 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
      661 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
      721 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
      781 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      841 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
      901 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
      961 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     1021 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     1081 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1141 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     1201 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1261 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     1321 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     1381 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1441 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     1501 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1561 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     1621 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     1681 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     1741 gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg
     1801 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     1861 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     1921 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     1981 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     2041 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     2101 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     2161 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     2221 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     2281 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     2341 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2401 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2461 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     2521 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     2581 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     2641 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     2701 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgctgccaag
     2761 cttatggaga aaaatcagtt tatcatcgat atttatgaaa agttgaatga gtaccacaag
     2821 aagctgctcg aactggcgag agcgctggca gaagcacttc gggccgggga tcgagcgcgc
     2881 gctggccgat tgctggagga gttccttgcg acgttcaaag caatgaaagc atacggagat
     2941 gctaaagagg tcgagagttt tctctacttc agtgaggagg aacttgcggc catatttgcg
     3001 gaatcagatg tcgaacggaa gaagctggga gtgacactca agatccctga agtcgtggcc
     3061 atggtagagg aactgggtcg cgcatttgag gctggggata ctgctactgc ggcggcgata
     3121 gtagaaaagt tggtaaagtt ttacgaggca aacataataa taggccaaaa gagaatcgat
     3181 gaactcaagt caaaaattgg cggtaccggc gggggtggct caggtggcat gccaggccca
     3241 accccatccg gcacaaatgt tgggtcaagc ggccgtagcc ctagtaaagc agtggccgca
     3301 cgcgccgccg gaagtaccgt gagacagcgc aaaaacgcat catgtggaac gcgctccgcc
     3361 gggaggacca cgtctgctgg gacagggggc atgtggaggt tctacaccga ggactctccg
     3421 ggactcaagg tgggtccagt tccggtgctc gtcatgtccc tgctgttcat tgcctccgtt
     3481 ttcatgcttc acatatgggg caaatacaca cgctcctgag gatccgcccc tctccctccc
     3541 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3601 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3661 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3721 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     3781 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3841 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3901 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3961 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     4021 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     4081 ctttgaaaaa cacgatgata atatggccga gctcatggcc ccaaagaaga agcggaaggt
     4141 cggtatccac ggagtcccag cagccatgag cgagctgatt aaggagaaca tgcacatgaa
     4201 gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg
     4261 caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc
     4321 cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca
     4381 cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca catgggagag
     4441 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
     4501 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt
     4561 gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg
     4621 cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc
     4681 aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt
     4741 ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga
     4801 gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct
     4861 taatggtgga ggtaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa
     4921 gggtggaggt ccaaaaaaga aaaggaaggt ctaagctagc ttgactgact gagatacagc
     4981 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     5041 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5101 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     5161 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     5221 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     5281 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     5341 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     5401 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     5461 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     5521 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     5581 accttgggct atcgaaactt aattaagtag gtgggcgggc caagataggg gcgcgattgc
     5641 tgcgatctgg aggacaaatt acacacactt gcgcctgagc gccaagcaca gggttgttgg
     5701 tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa tgttgccatg ggtagcatat
     5761 actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta gcataggcta
     5821 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     5881 tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     5941 tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta
     6001 tcctaataga gattagggta gtatatgcta tcctaattta tatctgggta gcatatacta
     6061 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat atgctatcct
     6121 aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     6181 aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct
     6241 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     6301 aatctgtatc cgggtagcat atgctatcct catgatatta attaaccagt caagtcagct
     6361 acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc
     6421 gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag
     6481 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6541 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6601 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6661 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6721 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6781 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6841 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6901 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6961 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7021 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7081 tttgcaagca gcagattacg cgcagaaaaa aaggatctca a
//
