LOCUS       Exported                6837 bp ds-DNA     circular SYN 27-APR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6837)
  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..6837
                     /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"
     primer_bind     895..909
                     /label=HR291_1_fwd_MfeI
     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"
     misc_feature    988..2355
                     /label=TeNT FL
     primer_bind     complement(2332..2358)
                     /label=HR291_1_rev
     promoter        complement(2389..2407)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2433..2657
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2703..3131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3145..3475
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3326..3461
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3542..4336
                     /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    4510..4631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4680..4696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4704..4720
                     /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(4728..4758)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4773..4794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5082..5670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5841..6701)
                     /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(6702..6806)
                     /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 caccggtccg atcaccatca acaacttccg ttactccgac
     1021 ccggttaaca acgacaccat catcatgatg gaaccgccgt actgcaaagg tctagacatc
     1081 tactacaaag ccttcaaaat caccgaccgt atctggatcg ttccggaacg ttacgaattt
     1141 ggtaccaaac cggaagactt caacccgccg tcctccctga tcgaaggtgc ttccgaatac
     1201 tacgacccga actacctgcg taccgactcc gacaaagacc gtttcctgca gaccatggtt
     1261 aaactgttca accgtatcaa aaacaacgtt gctggtgaag ctctgctgga caaaatcatc
     1321 aacgctatcc cgtacctggg taactcctac tccctgctgg acaaattcga caccaactcc
     1381 aactccgttt ccttcaacct gctggaacag gatccgtccg gtgctaccac caaatccgct
     1441 atgctgacca acctgatcat cttcggtccg ggtccggttc tgaacaaaaa cgaagttcgt
     1501 ggtatcgttc tgcgtgttga caacaaaaac tacttcccgt gccgtgacgg tttcggttcc
     1561 atcatgcaga tggctttctg cccggaatac gtaccgacct tcgacaacgt tatcgaaaac
     1621 atcacctccc tgaccatcgg taaatccaaa tacttccagg acccggctct gctgctgatg
     1681 cacgaactga tccacgttct gcacggtctg tacggcatgc aggtttcctc ccacgaaatc
     1741 atcccgtcca aacaggaaat ctacatgcag cacacctacc cgatctccgc tgaagagctc
     1801 ttcaccttcg gtggtcagga cgctaacctg atctccatcg atatcaaaaa cgacctgtac
     1861 gaaaaaaccc tgaacgacta caaagctatc gctaacaaac tgtcccaggt tacctcctgc
     1921 aacgacccga acatcgacat cgactcctac aaacagatct atcagcagaa gtatcagttt
     1981 gataaggatt ccaacggcca gtatatagtg aacgaggata agtttcagat actgtataac
     2041 tccataatgt atggctttac tgagatagag ctgggcaaga agtttaacat aaagactcgc
     2101 ttaagctatt tttccatgaa ccacgatcct gtgaagatac ctaacctgct ggatgatact
     2161 atatataacg atactgaggg ctttaacata gagtccaagg atctgaagtc cgagtataag
     2221 ggccagaaca tgagggtgaa cactaacgcc tttaggaacg tcgacggttc cggtctggtt
     2281 tccaaactga tcggtctgtg caaaaaaatc atcccgccga ccaacatccg tgaaaacctg
     2341 tacaaccgta ccgcttaata actcgagcat gcatctagag ggccctattc tatagtgtca
     2401 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2461 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2521 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2581 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2641 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2701 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2761 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2821 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     2881 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2941 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3001 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3061 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3121 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3181 ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3481 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3541 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3601 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3661 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3721 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3781 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3841 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3901 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3961 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4021 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4081 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4141 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4201 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4261 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4321 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4381 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4441 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4501 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4561 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4621 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4741 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4861 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     5281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5521 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
