LOCUS       Exported                6677 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 6677)
  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..6677
                     /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"
     primer_bind     906..924
                     /label=Smbit-sbf-f
                     /note="tm 56"
     CDS             907..942
                     /codon_start=1
                     /label=SmBiT
                     /translation="MVTGYRLFEEIL"
     primer_bind     907..924
                     /label=SmBit-GSSG-sbf-f
                     /note="Tm 54"
     primer_bind     complement(924..943)
                     /label=Smbit-Cla-r
                     /note="Tm 54"
     CDS             943..975
                     /codon_start=1
                     /label=Linker
                     /translation="SGLKSRRALDS"
     CDS             976..2181
                     /codon_start=1
                     /label=New Arr3
                     /translation="MGEKPGTRVFKKSSPNCKLTVYLGKRDFVDHLDKVDPVDGVVLVD
                     PDYLKDRKVFVTLTCAFRYGREDLDVLGLSFRKDLFIANYQAFPPTPNPPRPPTRLQER
                     LLRKLGQHAHPFFFTIPQNLPCSVTLQPGPEDTGKACGVDFEIRAFCAKSLEEKSHKRN
                     SVRLVIRKVQFPPGPETTRHFLMSDRSLHLEASLDKELYYHGEPLNVNVHVTNNSTKTV
                     KKIKVSVRQYADICLFSTAQYKCPVAQVEQDDQVSPSSTFCKVYTITPLLSNNREKRGL
                     ALDGKLKHEDTNLASSTIVKEGANKEVLGILVSYRVKVKLVVSRGGDVSVELPFVLMHP
                     KPHDHIALPRPQSAVPETDAPVDTNLIEFETNYATDDDIVFEDFARLRLKGLKDEDYDD
                     QFC"
     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..1578
                     /codon_start=1
                     /label=T3 V172-L208
                     /translation="VQFPPGPETTRHFLMSDRSLHLEASLDKEL"
     mat_peptide     1498..1509
                     /product="175 APEKPGPQPSAto
                     175 PPGP
                     deletion 175,177-178, 182-185
                     "
                     /label=delta7
     CDS             1498..1509
                     /codon_start=1
                     /label=T3 V172-L208
                     /translation="PPGP"
     primer_bind     1568..1602
                     /label=TNCO
     CDS             1579..1632
                     /codon_start=1
                     /label=T4 Y209-T226
                     /translation="YYHGEPLNVNVHVTNNST"
     CDS             1750..1812
                     /codon_start=1
                     /label=T5 S226-R286
                     /translation="SSTFCKVYTITPLLSNNREKR"
     primer_bind     1811..1838
                     /label=AT10f
     CDS             1813..1908
                     /codon_start=1
                     /label=T6 G287-I318
                     /translation="GLALDGKLKHEDTNLASSTIVKEGANKEVLGI"
     CDS             1987..2181
                     /codon_start=1
                     /label=T7 M345-C409
                     /translation="MHPKPHDHIALPRPQSAVPETDAPVDTNLIEFETNYATDDDIVFE
                     DFARLRLKGLKDEDYDDQFC"
     primer_bind     complement(2116..2133)
                     /label=A3Tr393-Cla-r
     CDS             2188..2211
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2229..2247)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2273..2497
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2367..2386)
                     /label=P3BHGR
                     /note="BGH Revese, sequencing primer"
     rep_origin      2543..2971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2985..3315
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3166..3301
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3382..4176
                     /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    4350..4471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4520..4536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4922..5510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5681..6541)
                     /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(6542..6646)
                     /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 gatcctgtag atggtgtggt gctagtggac ccggactact tgaaggaccg caaagtgttt
     1141 gtgaccctca cctgcgcctt tcgctatggc cgcgaagacc tggacgtgct gggcttgtcc
     1201 ttccgcaaag acctgttcat cgccaactac caggccttcc ccccaacacc caacccaccc
     1261 cggcccccca cccgcctgca ggagcggctg ctgaggaagc tgggccagca tgcccacccc
     1321 ttttttttca caatacccca gaacctgccc tgctccgtca ccctgcagcc aggcccagag
     1381 gatacgggga aggcctgcgg ggtcgacttt gagattcgag ccttctgcgc caaatcacta
     1441 gaagagaaaa gccacaaacg gaactctgtg cgtctggtga tccgaaaggt gcagtttcca
     1501 cccggccccg aaaccacacg ccacttcctc atgtctgacc ggtccctgca ccttgaggct
     1561 tcactggaca aggagctgta ctaccacggg gagcccctca atgtcaatgt ccacgtcacc
     1621 aataactcca ccaagactgt caagaagatc aaagtctctg tgagacagta cgccgacatc
     1681 tgcctcttca gcaccgccca gtacaagtgc cccgtggctc aggtcgaaca agatgaccag
     1741 gtgtcgccca gttccacgtt ctgtaaggtg tacaccataa ccccgctgct cagcaacaac
     1801 cgggagaagc gcggcctcgc tctagatggg aagctcaagc acgaggacac caacctggcc
     1861 tcgagcacca ttgtgaagga gggtgccaac aaggaggtgc tggggatact ggtgtcctac
     1921 agggtcaagg tgaagctggt ggtgtcacga ggcggggatg tctcagtgga gctgccattt
     1981 gttctcatgc accccaagcc ccatgatcac atcgcccttc ccaggcccca gtcagctgtt
     2041 cctgaaacag atgcccccgt ggacaccaac ctcattgaat ttgaaaccaa ctatgccaca
     2101 gacgacgaca tcgtgttcga agactttgcc cggctgcggc tcaaggggct gaaggacgag
     2161 gactatgacg accagttctg tatcgatgat tacaaggatg acgacgataa gtagaagctt
     2221 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     2281 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2341 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2401 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2461 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2521 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2581 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2641 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     2701 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2761 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2821 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2881 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     2941 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3001 gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag
     3061 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3181 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3241 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3301 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3361 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3421 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3481 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3541 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3601 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3661 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3721 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3781 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3841 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3901 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3961 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4021 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4081 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4141 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4201 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4261 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4321 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4381 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4441 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4501 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4561 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4621 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4681 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4741 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4801 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4861 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4921 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4981 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5041 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5101 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5161 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5221 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5281 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5341 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5401 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5461 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5521 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5581 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5641 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5701 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5761 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5821 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5881 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5941 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6001 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6061 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6121 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6181 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6241 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6301 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6361 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6421 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6481 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6541 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6601 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6661 aagtgccacc tgacgtc
//
