LOCUS       Exported                8519 bp ds-DNA     circular SYN 06-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8519)
  AUTHORS   Heegwang Roh
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 9, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8519
                     /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"
     CDS             4011..4040
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(4071..4089)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    4115..4339
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4385..4813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4827..5157
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5008..5143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5224..6018
                     /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    6192..6313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6362..6378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6386..6402
                     /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(6410..6440)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6455..6476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6764..7352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7523..8383)
                     /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(8384..8488)
                     /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 gcatgccgat caccatcaac aacttccgtt actccgaccc ggttaacaac gacaccatca
      961 tcatgatgga accgccgtac tgcaaaggtc tagacatcta ctacagagcc ttcaaaatca
     1021 ccgaccgtat ctggatcgtt ccggaacgtt acgaatttgg taccaaaccg gaagacttca
     1081 acccgccgtc ctccctgatc gaaggtgctt ccgaatacta cgacccgaac tacctgcgta
     1141 ccgactccga caaagaccgt ttcctgcaga ccatggttaa actgttcaac cgtatcaaaa
     1201 acaacgttgc tggtgaagct ctgctggaca aaatcatcaa cgctatcccg tacctgggta
     1261 actcctactc cctgctggac aaattcgaca ccaactccaa ctccgtttcc ttcaacctgc
     1321 tggaacagga tccgtccggt gctaccacca aatccgctat gctgaccaac ctgatcatct
     1381 tcggtccggg tccggttctg aacaaaaacg aagttcgtgg tatcgttctg cgtgttgaca
     1441 acaaaaacta cttcccgtgc cgtgacggtt tcggttccat catgcagatg gctttctgcc
     1501 cggaatacgt accgaccttc gacaacgtta tcgaaaacat cacctccctg accatcggta
     1561 aatccaaata cttccaggac ccggctctgc tgctgatgca cgaactgatc cacgttctgc
     1621 acggtctgta cggcatgcag gtttcctccc acgaaatcat cccgtccaaa caggaaatct
     1681 acatgcagca cacctacccg atctccgctg aagagctctt caccttcggt ggtcaggacg
     1741 ctaacctgat ctccatcgat atcaaaaacg acctgtacga aaaaaccctg aacgactaca
     1801 aagctatcgc taacaaactg tcccaggtta cctcctgcaa cgacccgaac atcgacatcg
     1861 actcctacaa acagatctat cagcagaagt atcagtttga taaggattcc aacggccagt
     1921 atatagtgaa cgaggataag tttcagatac tgtataactc cataatgtat ggcgggagcg
     1981 gcagcggggg gtccggtagc ggggagtttc gggcaaccac actggaacgg atcgagaaaa
     2041 gcttcctgat tactgatccg agactgcctg acaacccaat catttttgtg agcaattcct
     2101 tcctgcagct gacagaatat tctcgggaag agatcctggg gcgcaattgc cgttttctgc
     2161 agggacccga gacagaccgt gccactgttc ggaaaatcag agatgctatt gacaaccaga
     2221 ctgaagtgac cgttcagctg atcaattata ccaagagcgg caagaagttc tggaacgtgt
     2281 tccacctgca gccgatgcgc gattataagg gcgacgtcca gtacttcatt ggcgtgcagc
     2341 tggatggcac cgaacgcctt catggcgccg ctgagcgtga ggcggtctgc ctggtcaaaa
     2401 agacagcctt tcagattttt actgagatag agctgggcaa gaagtttaac ataaagactc
     2461 gcttaagcta tttttccatg aaccacgatc ctgtgaagat acctaacctg ctggatgata
     2521 ctatatataa cgatactgag ggctttaaca tagagtccaa ggatctgaag tccgagtata
     2581 agggccagaa catgagggtg aacactaacg cctttaggaa cgtcgacggt tccggtctgg
     2641 tttccaaact gatcggtctg tgcaaaaaaa tcatcccgcc gaccaacatc cgtgaaaacc
     2701 tgtacaaccg taccgctggc tcgggcggga gtaccggtgg tagtgggagc ggtggtgaaa
     2761 aacccgggac cagggtcttc aagaagtcga gccctaactg caagctcacc gtgtacttgg
     2821 gcaagcgtga ctttgtggat cacttggaca aagtggatcc tgtcgatggt gtggtgcttg
     2881 tggatcctga ctacttgaag gaccggaaag tgtttgtgac cctcacctgt gccttccgct
     2941 atggccgaga agacctggat gtactgggcc tgtctttccg caaagatctg ttcatcgcca
     3001 cctaccaggc cttccccccc atgcccaacc cacctcggcc ccccacccgc ctacaggacc
     3061 gactgctgaa gaagttgggc cagcacgccc accccttttt tttcacaata ccccagaatt
     3121 tgccttgctc cgtcacactg cagccaggac cggaggacac agggaaggcc tgtggagtag
     3181 actttgagat tcgagccttc tgtgccaaat ctatagaaga aaaaagccac aaaaggaact
     3241 ccgtgcggct tatcatcaga aaggtacagt ttgctcctga gacacccggc ccccagccat
     3301 cagctgaaac cacacgccac ttcctcatgt ctgaccggag gtccctgcac ctagaggctt
     3361 ccctggacaa agagctgtac taccatgggg aacccctcaa tgtcaacgtc cacgtcacca
     3421 acaattctgc caaaaccgtc aagaagatca gagtgtctgt gagacagtat gccgacattt
     3481 gcctcttcag caccgcgcag tacaagtgtc ctgtggctca gcttgaacaa gatgaccagg
     3541 tgtctcccag ttccacattc tgcaaggtgt acaccataac cccgctgctc agtgacaacc
     3601 gagagaagcg tggccttgcc cttgatgggc aactcaagca cgaagacacc aacctggctt
     3661 ccagcaccat tgtgaaggag ggagccaaca aggaggtgct gggaatccta gtatcctaca
     3721 gggtcaaggt gaagctggtg gtgtctcgag gcggggatgt ctccgtggag ctacctttcg
     3781 tcctaatgca ccccaagccc cacgaccaca tcacccttcc ccgaccccag tcagcccccc
     3841 gggaaataga catccctgtg gataccaacc tcattgaatt cgataccaac tatgccacag
     3901 acgacgacat cgtgtttgag gactttgcga ggcttcggct gaaggggatg aaggatgacg
     3961 actgtgatga ccagttctgc ggtggaagtg gatcaggcag cggtggatct gaacaaaagc
     4021 ttatttctga agaggacttg taactcgagc atgcatctag agggccctat tctatagtgt
     4081 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     4141 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4201 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4261 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4321 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4381 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     4561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4741 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4801 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4861 ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4921 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4981 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5041 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5101 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5161 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5221 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5281 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5341 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5401 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5461 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5521 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5581 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5641 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5701 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5761 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5821 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5881 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5941 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6001 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6061 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6121 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6181 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6241 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6301 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6361 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6421 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6481 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6541 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6601 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6661 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6721 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6781 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6841 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6901 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     6961 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7021 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7081 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7141 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7201 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7261 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7321 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7381 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7441 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7501 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7561 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7621 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7681 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7741 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7801 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7861 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7921 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7981 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8041 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8101 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8161 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8221 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8281 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8341 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8401 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8461 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
