LOCUS       Exported                6083 bp ds-DNA     circular SYN 10-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  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..6083
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        206..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        758..776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     796..855
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
     misc_feature    856..1230
                     /label=GFPnb
     misc_feature    1261..1305
                     /label=ALFAtag
     misc_feature    1336..2475
                     /label=KORD
     misc_feature    2476..2511
                     /label=NES
     CDS             2524..3033
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             3034..3075
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3115..3322
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3351..3491
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3566..4021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4303..4407
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4408..5268
                     /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"
     rep_origin      5439..6027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtt gtacatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      721 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      781 cccaagcttg ccaccatgta caggatgcaa ctcctgtctt gcattgcact aagtcttgca
      841 cttgtcacga attcggtgca actcgtagag tccggtggcg gactcgtcca ggcgggaggc
      901 tccctgcgcc tgagttgcgc tgcgtccgga agaactatta gcatggccgc catgagttgg
      961 ttcagacagg cgcctggaaa ggagagggag ttcgtggcag gcataagtcg ctccgcaggg
     1021 tcagccgtcc atgccgactc cgtaaaggga cgctttacca tctccagaga taacacaaaa
     1081 aacacgctct atctccaaat gaattctttg aaggccgagg acacagccgt gtactactgc
     1141 gccgttagga ccagcggatt ttttggaagc attcctagga cagggacagc ctttgattac
     1201 tggggccagg gaacgcaggt taccgtttcc accggtggtg gcggctccgg tggagggtct
     1261 ccgtccagac ttgaggaaga gttgagacgg aggttgaccg agccaggcgg tggcagtgga
     1321 ggtgggtccg gtaccatgga aagccccatt caaatcttcc ggggcgagcc cggaccaacc
     1381 tgcgctccaa gcgcttgcct gccaccgaat tctagtgcat ggttcccagg gtgggcagag
     1441 cccgatagca atggctcagc cgggtccgag gatgcgcagc ttgaacccgc tcacatctca
     1501 cccgctatcc ctgtcattat tacggcagtc tactcagttg ttttcgttgt gggcctggtt
     1561 ggaaatagcc tcgtgatgtt tgtgattatc aggtatacta aaatgaagac cgccacgaat
     1621 atctacatct ttaacctggc tctggctgac gccttggtga ccaccactat gccatttcag
     1681 tccaccgtct atcttatgaa ctcttggcca ttcggcgacg ttctgtgcaa gattgtcatt
     1741 tccattaatt attataatat gtttacatca atctttacgc ttacaatgat gagcgtggac
     1801 cgatacattg ctgtttgcca tccggtgaaa gcactggatt ttcgcacccc cttgaaggcc
     1861 aagattatca atatctgtat ttggctcctg tcctcatccg tcggaatttc cgccattgtc
     1921 ttgggtggta caaaggtgag ggaggacgtg gacgttatag agtgcagcct gcagttccct
     1981 gacgatgact acagctggtg ggaccttttt atgaagatct gtgtcttcat ttttgctttc
     2041 gtcatccccg tgctgattat catagtgtgc tatacactga tgatattgcg gctgaaatct
     2101 gtgcgcctgc tgagcggcag ccgcgaaaag gatcggaatc tgcgccgaat cacacgcctc
     2161 gtgctggttg tagtcgctgt cttcgtggta tgctggaccc caatccacat tttcatcctg
     2221 gtggaagctc tgggatccac ctctcactcc actgccgcac ttagctcata ttacttctgc
     2281 attgccttgg gctacactaa ttcctcactg aatcccattc tgtacgcatt tcttgacgag
     2341 aacttcaaac ggtgctttag agacttttgc ttccccctca agatgcgcat ggagcggcag
     2401 tcaaccagcc gagtccggaa tacggtccag gatcccgcct atctgcggga catcgacggc
     2461 atgaacaagc ctgtggagct ggccgagaag ctggccggcc tggacatcaa cggtggagct
     2521 agcgtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg
     2581 gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta
     2641 actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc
     2701 atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag
     2761 gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta
     2821 atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc
     2881 gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc
     2941 gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg
     3001 accggctggc ggctgtgcga acgcattctg gcgggcaagc cgatccctaa ccctctgctg
     3061 ggcctggaca gcacttagga attcctcgag ctagagctcg ctgatcagcc tcgactgtgc
     3121 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3181 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3241 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3301 acaatagcag gcatgctggg gaggtaacca cgtgcggacc gagcggccgc aggaacccct
     3361 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3421 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3481 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3541 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3601 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     3661 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3721 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3781 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3841 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     3901 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     3961 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4021 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4081 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4141 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4201 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4261 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4321 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4381 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4441 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4501 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4561 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4621 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4681 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4741 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4801 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4861 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4921 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4981 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5041 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5101 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
     5161 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5221 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5281 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5341 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5401 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5461 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5521 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5581 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5641 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5701 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5761 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5821 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5881 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5941 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6001 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6061 gccttttgct ggccttttgc tca
//
