LOCUS       Exported                7821 bp ds-DNA     circular SYN 11-JAN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7821)
  AUTHORS   Heegwang Roh
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 6, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7821
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             910..951
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             988..1497
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     misc_feature    1528..2595
                     /label=TeNT N LATeNT2
     misc_feature    1628
                     /label=A>G (K34R)
     misc_feature    2626..3039
                     /label=hLOV(-1) LATeNT2
     misc_feature    2716
                     /label=G>A (D397N)
     misc_feature    2979
                     /label=T>C (silent)
     misc_feature    3040..3339
                     /label=TeNT C
     promoter        complement(3373..3391)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3417..3641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3687..4115
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4129..4459
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4310..4445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4526..5320
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5494..5615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5664..5680)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5688..5704
                     /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)."
     promoter        complement(5712..5742)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5757..5778
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6066..6654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6825..7685)
                     /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"
     promoter        complement(7686..7790)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc
      901 gccaccatgg gcaagcccat ccccaacccc ctgctgggcc tggacagcac caagggctcg
      961 ggctcgacct cgggctcggg caccggtgtc ttcacactcg aagatttcgt tggggactgg
     1021 cgacagacag ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg
     1081 tttcagaatc tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat
     1141 gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg
     1201 ggccagatcg aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg
     1261 atcctgcact atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc
     1321 ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc
     1381 ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg
     1441 ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcgggg
     1501 agcggcagcg gggggtccgg tagcgggccg atcaccatca acaacttccg ttactccgac
     1561 ccggttaaca acgacaccat catcatgatg gaaccgccgt actgcaaagg tctagacatc
     1621 tactacagag ccttcaaaat caccgaccgt atctggatcg ttccggaacg ttacgaattt
     1681 ggtaccaaac cggaagactt caacccgccg tcctccctga tcgaaggtgc ttccgaatac
     1741 tacgacccga actacctgcg taccgactcc gacaaagacc gtttcctgca gaccatggtt
     1801 aaactgttca accgtatcaa aaacaacgtt gctggtgaag ctctgctgga caaaatcatc
     1861 aacgctatcc cgtacctggg taactcctac tccctgctgg acaaattcga caccaactcc
     1921 aactccgttt ccttcaacct gctggaacag gatccgtccg gtgctaccac caaatccgct
     1981 atgctgacca acctgatcat cttcggtccg ggtccggttc tgaacaaaaa cgaagttcgt
     2041 ggtatcgttc tgcgtgttga caacaaaaac tacttcccgt gccgtgacgg tttcggttcc
     2101 atcatgcaga tggctttctg cccggaatac gtaccgacct tcgacaacgt tatcgaaaac
     2161 atcacctccc tgaccatcgg taaatccaaa tacttccagg acccggctct gctgctgatg
     2221 cacgaactga tccacgttct gcacggtctg tacggcatgc aggtttcctc ccacgaaatc
     2281 atcccgtcca aacaggaaat ctacatgcag cacacctacc cgatctccgc tgaagagctc
     2341 ttcaccttcg gtggtcagga cgctaacctg atctccatcg atatcaaaaa cgacctgtac
     2401 gaaaaaaccc tgaacgacta caaagctatc gctaacaaac tgtcccaggt tacctcctgc
     2461 aacgacccga acatcgacat cgactcctac aaacagatct atcagcagaa gtatcagttt
     2521 gataaggatt ccaacggcca gtatatagtg aacgaggata agtttcagat actgtataac
     2581 tccataatgt atggcgggag cggcagcggg gggtccggta gcggggagtt tcgggcaacc
     2641 acactggaac ggatcgagaa aagcttcgtg attactgatc cgagactgcc tgacaaccca
     2701 atcatttttg tgagcaattc cttcctgcag ctgacagaat attctcggga agagatcctg
     2761 gggcgcaatt gccgttttct gcagggaccc gagacagacc gtgccactgt tcggaaaatc
     2821 agagatgcta ttgacaacca gactgaagtg accgttcagc tgatcaatta taccaagagc
     2881 ggcaagaagt tctggaacgt gttccacctg cagccgatgc gcgattataa gggcgacgtc
     2941 cagtacttca ttggcgtgca gctggatggc accgaacgcc ttcatggcgc cgctgagcgt
     3001 gaggcggtct gcctggtcaa aaagacagcc tttcagattt ttactgagat agagctgggc
     3061 aagaagttta acataaagac tcgcttaagc tatttttcca tgaaccacga tcctgtgaag
     3121 atacctaacc tgctggatga tactatatat aacgatactg agggctttaa catagagtcc
     3181 aaggatctga agtccgagta taagggccag aacatgaggg tgaacactaa cgcctttagg
     3241 aacgtcgacg gttccggtct ggtttccaaa ctgatcggtc tgtgcaaaaa aatcatcccg
     3301 ccgaccaaca tccgtgaaaa cctgtacaac cgtaccgctt aataactcga gcatgcatct
     3361 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     3421 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3481 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3541 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3601 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3661 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3721 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3781 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3841 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3901 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3961 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4021 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4081 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4141 gggtgtggaa agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa
     4201 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4261 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4321 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4381 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4441 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4501 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4561 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4621 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4681 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4741 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4801 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4861 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4921 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4981 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5041 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5101 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5161 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5221 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5281 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5341 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5401 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5461 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5521 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5581 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5641 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5701 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5761 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5821 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5881 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5941 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6001 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6061 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6241 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6481 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6781 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6841 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6901 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6961 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7021 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7081 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7141 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7201 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7261 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7321 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7381 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7441 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7501 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7561 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7621 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7681 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7741 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7801 cgaaaagtgc cacctgacgt c
//
