LOCUS       sequence-424650-        6246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6246)
  AUTHORS   Tang WF, Yang SY, Wu BW, Jheng JR, Chen YL, Shih CH, Lin KH, Lai HC,
            Tang P, Horng JT
  TITLE     Reticulon 3 binds the 2C protein of enterovirus 71 and is required 
            for viral replication.
  JOURNAL   J Biol Chem. 2007 Feb 23. 282(8):5888-98.
  PUBMED    17182608
REFERENCE   2  (bases 1 to 6246)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2007 Feb 23. 282(8):5888-98."
COMMENT     Sequence Label: pcDNA3.1-Myc-His RTN3A1
FEATURES             Location/Qualifiers
     source          1..6246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..55)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        40..369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      220..355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     282..301
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             436..1230
                     /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"
     primer_bind     complement(490..509)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1100..1119
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1404..1525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1441..1460)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1495..1514
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1574..1590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1574..1590)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1587..1609)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1598..1614
                     /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(join(1622..1628,1629..1646,1647..1652))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1667..1688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1805..1822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1976..2564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2056..2075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2735..3526,3527..3595))
                     /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"
     primer_bind     3358..3377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3596..3700)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3775..3794)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3966..4345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4346..4549
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4500..4520
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4594..4613
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4594..4612
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5462..5491
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5507..5524
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5568..5585)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5574..5798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5844..6246,1..26)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5931..5950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6141..6162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
       61 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
      121 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
      181 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
      241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
      301 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
      361 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
      421 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
      481 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
      541 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
      601 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
      661 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
      721 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
      781 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
      841 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
      901 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
      961 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     1021 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     1081 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     1141 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     1201 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     1261 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     1321 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     1381 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     1441 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1501 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     1561 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1621 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1681 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1741 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1801 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1861 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1921 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1981 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2041 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2101 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2161 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2221 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2281 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2341 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2401 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2461 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2521 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2581 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2641 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2701 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2761 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2821 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2881 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2941 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3001 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3061 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3121 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3181 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3241 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3301 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3361 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3421 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3481 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3541 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3601 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3661 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3721 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     3781 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     3841 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     3901 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     3961 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     4021 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     4081 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     4141 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     4201 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     4261 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     4321 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     4381 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     4441 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     4501 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
     4561 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
     4621 agctggctag cgtttaaacg ggccctctag actcgagcgg ccgccactgt gctggatatc
     4681 tgcagaattc gaccatatgg cggagccgtc ggcggccact cagtcccatt ccatctcctc
     4741 gtcgtccttc ggagccgagc cgtccgcgcc cggcggcggc gggagcccag gagcctgccc
     4801 cgccctgggg acgaagagct gcagctcctc ctgtgcggtg cacgatctga ttttctggag
     4861 agatgtgaag aagactgggt ttgtctttgg caccacgctg atcatgctgc tttccctggc
     4921 agctttcagt gtcatcagtg tggtttctta cctcatcctg gctcttctct ctgtcaccat
     4981 cagcttcagg atctacaagt ccgtcatcca agctgtacag aagtcagaag aaggccatcc
     5041 attcaaagcc tacctggacg tagacattac tctgtcctca gaagctttcc ataattacat
     5101 gaatgctgcc atggtgcaca tcaacagggc cctgaaactc attattcgtc tctttctggt
     5161 agaagatctg gttgactcct tgaagctggc tgtcttcatg tggctgatga cctatgttgg
     5221 tgctgttttt aacggaatca cccttctaat tcttgctgaa ctgctcattt tcagtgtccc
     5281 gattgtctat gagaagtaca agacccagat tgatcactat gttggcatcg cccgagatca
     5341 gaccaagtca attgttgaaa agatccaagc aaaactccct ggaatcgcca aaaaaaaggc
     5401 agaaagggtc gaattccacc acactggact agtggatccg agctcggtac caagctttct
     5461 agaacaaaaa ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca
     5521 tcattgagtt taaacggtct ccagcttaag tttaaaccgc tgatcagcct cgactgtgcc
     5581 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     5641 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     5701 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     5761 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     5821 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5881 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5941 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6001 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6061 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6121 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6181 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6241 tgagct
//
