LOCUS       Exported                6650 bp ds-DNA     circular SYN 02-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6650)
  AUTHORS   Chen Zheng
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 13, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6650
                     /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"
     primer_bind     769..789
                     /label=CMVf
                     /note="Sequencing primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             907..942
                     /codon_start=1
                     /label=SmBiT
                     /translation="MVTGYRLFEEIL"
     CDS             943..975
                     /codon_start=1
                     /label=Linker
                     /translation="SGLKSRRALDS"
     CDS             976..2154
                     /codon_start=1
                     /label=New Arr3 1-393
                     /translation="MGEKPGTRVFKKSSPNCKLTVYLGKRDFVDHLDKVDPVDGVVLVD
                     PDYLKDRKVFVTLTCAFRYGREDLDVLGLSFRKDLFIANYQAFPPTPNPPRPPTRLQER
                     LLRKLGQHAHPFFFTIPQNLPCSVTLQPGPEDTGKACGVDFEIRAFCAKSLEEKSHKRN
                     SVRLVIRKVQFAPEKPGPQPSAETTRHFLMSDRSLHLEASLDKELYYHGEPLNVNVHVT
                     NNSTKTVKKIKVSVRQYADICLFSTAQYKCPVAQVEQDDQVSPSSTFCKVYTITPLLSN
                     NREKRGLALDGKLKHEDTNLASSTIVKEGANKEVLGILVSYRVKVKLVVSRGGDVSVEL
                     PFVLMHPKPHDHIALPRPQSAVPETDAPVDTNLIEFETNYATDDDIVFEDFAR"
     CDS             976..1131
                     /codon_start=1
                     /label=T1
                     /translation="MGEKPGTRVFKKSSPNCKLTVYLGKRDFVDHLDKVDPVDGVVLVD
                     PDYLKDR"
     primer_bind     1068..1105
                     /label=AT2f
     CDS             1234..1302
                     /codon_start=1
                     /label=T2A
                     /translation="AFPPTPNPPRPPTRLQERLLRKL"
     CDS             1303..1347
                     /codon_start=1
                     /label=T2B G110-L124
                     /translation="GQHAHPFFFTIPQNL"
     CDS             1489..1599
                     /codon_start=1
                     /label=T3 V172-L208
                     /translation="VQFAPEKPGPQPSAETTRHFLMSDRSLHLEASLDKEL"
     primer_bind     1589..1623
                     /label=TNCO
     CDS             1600..1653
                     /codon_start=1
                     /label=T4 Y209-T226
                     /translation="YYHGEPLNVNVHVTNNST"
     CDS             1771..1833
                     /codon_start=1
                     /label=T5 S226-R286
                     /translation="SSTFCKVYTITPLLSNNREKR"
     primer_bind     1832..1859
                     /label=AT10f
     CDS             1834..1929
                     /codon_start=1
                     /label=T6 G287-I318
                     /translation="GLALDGKLKHEDTNLASSTIVKEGANKEVLGI"
     CDS             2008..2154
                     /codon_start=1
                     /label=T7 M345-C409
                     /translation="MHPKPHDHIALPRPQSAVPETDAPVDTNLIEFETNYATDDDIVFE
                     DFAR"
     primer_bind     complement(2137..2163)
                     /label=A3Tr393-Cla-r
     CDS             2161..2184
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2202..2220)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2246..2470
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2340..2359)
                     /label=P3BHGR
                     /note="BGH Revese, sequencing primer"
     rep_origin      2516..2944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2958..3288
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3139..3274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3355..4149
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4323..4444
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4493..4509)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4895..5483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5654..6514)
                     /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(6515..6619)
                     /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 gctggaattc
      901 accaccatgg tgaccggcta ccggctgttc gaggagattc tctccggact taagtctcgg
      961 cgagcgctcg actccatggg ggagaaaccc gggaccaggg tcttcaagaa atcgagtcct
     1021 aactgcaagc tcaccgtgta cctaggcaag cgggactttg tggaccacct ggacaaagtg
     1081 gaccctgtag atggtgtggt gctagtggat ccggactact tgaaggaccg caaagtgttt
     1141 gtgaccctca cgtgcgcctt tcgctatggc cgcgaagacc tggacgtcct gggcttgtcc
     1201 ttccgcaaag acctgttcat cgccaactac caggccttcc ctccaacacc taacccacct
     1261 agaccaccta cgcgtctgca ggagcggctg ctgaggaagc tgggccagca tgcccacccc
     1321 ttttttttca caatacccca gaacctgccc tgctccgtca ccctacagcc aggcccagag
     1381 gatacgggga aggcttgcgg ggtcgacttt gagattcgag ccttctgcgc caaatcacta
     1441 gaagagaaaa gccacaaacg gaactccgta cgtctggtga tccgaaaggt gcagtttgcc
     1501 ccagagaaac cagggcccca gccttcagct gaaaccacac gccacttcct catgtctgac
     1561 cggtccctgc accttgaggc ttcactggac aaggagctgt actaccacgg ggagcccctc
     1621 aatgtcaatg tccacgtcac caataactcc accaagactg tcaagaagat caaagtctct
     1681 gtgagacagt acgccgacat ctgcctcttc agcaccgccc agtacaagtg ccccgtggct
     1741 caggtcgaac aagatgacca ggtgtcgccc agttccacgt tctgtaaggt gtacaccata
     1801 accccgctgc tcagcaacaa ccgggagaag cgcggcctcg ctctagatgg gaagctcaag
     1861 cacgaggaca ccaacctggc ctcgagcacc attgtgaagg agggtgccaa caaggaggtg
     1921 ctggggatac tggtgtccta cagggtcaag gtgaagctgg tggtgtcacg aggcggggat
     1981 gtctcagtgg agctgccatt tgttctcatg caccccaagc cccatgatca catcgccctt
     2041 cccaggcccc agtcagctgt tcctgaaaca gatgcccccg tggacaccaa cctcattgaa
     2101 tttgaaacca actatgccac agacgacgac atcgtgttcg aagactttgc ccggatcgat
     2161 gattacaagg atgacgacga taagtagaag cttggcccta ttctatagtg tcacctaaat
     2221 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2281 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2341 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2401 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2461 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2641 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     2701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2881 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2941 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccaggca
     3001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3061 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3121 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3181 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3241 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3301 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3361 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3421 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3481 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3541 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3601 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3661 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3721 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3781 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3841 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3901 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3961 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4021 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4081 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4141 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4201 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4261 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4321 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4381 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4441 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4501 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4561 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4621 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4681 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4741 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4801 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4861 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4921 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4981 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5041 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     5101 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5161 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5221 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5281 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5341 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5401 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5641 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5701 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5761 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5821 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5881 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5941 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6001 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6061 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6121 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6181 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6241 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6301 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6361 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6421 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6481 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6541 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6601 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
