LOCUS       sequence-372910-        6164 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 6164)
  AUTHORS   Wilker EW, Grant RA, Artim SC, Yaffe MB
  TITLE     A structural basis for 14-3-3sigma functional specificity.
  JOURNAL   J Biol Chem. 2005 May 13. 280(19):18891-8.
  PUBMED    15731107
REFERENCE   2  (bases 1 to 6164)
  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. 2005 May 13. 280(19):18891-8."
COMMENT     Sequence Label: pcDNA3 HA 14-3-3 sigma
FEATURES             Location/Qualifiers
     source          1..6164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..44)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        29..358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      209..344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     271..290
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             425..1219
                     /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(479..498)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1089..1108
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1393..1514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1430..1449)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1484..1503
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1563..1579)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1563..1579)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1576..1598)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1587..1603
                     /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(1611..1617,1618..1635,1636..1641))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1656..1677
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1794..1811)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1965..2553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2045..2064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2724..3515,3516..3584))
                     /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     3347..3366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3585..3689)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3764..3783)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3955..4334
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4335..4538
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4489..4509
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4583..4602
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4583..4601
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4624..4650
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             4624..4650
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4630..4656
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(5437..5455)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5438..5455)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5475..5492)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5481..5705
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5751..6164,1..15)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5838..5857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6048..6069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
       61 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      121 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      181 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      241 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      301 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      361 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
      421 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
      481 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
      541 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
      601 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
      661 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
      721 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
      781 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
      841 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
      901 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
      961 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1021 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1081 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1141 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     1201 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     1261 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     1321 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     1381 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1441 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1501 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     1561 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1621 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1681 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1741 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1801 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1861 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1921 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1981 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2041 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2101 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2161 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2221 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2281 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2341 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2401 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2461 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2521 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2581 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2641 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2701 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2761 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2821 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2881 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2941 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3001 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3061 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3121 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3181 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3241 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3301 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3361 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3421 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3481 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3541 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3601 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3661 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3721 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     3781 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     3841 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     3901 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     3961 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     4021 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4081 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4141 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4201 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4261 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4321 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4381 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4441 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4501 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
     4561 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg
     4621 gcctacccct acgacgtgcc cgactacgcc tccctcggat ccatggagag agccagtctg
     4681 atccagaagg ccaagctggc agagcaggcc gaacgctatg aggacatggc agccttcatg
     4741 aaaggcgccg tggagaaggg cgaggagctc tcctgcgaag agcgaaacct gctctcagta
     4801 gcctataaga acgtggtggg cggccagagg gctgcctgga gggtgctgtc cagtattgag
     4861 cagaaaagca acgaggaggg ctcggaggag aaggggcccg aggtgcgtga gtaccgggag
     4921 aaggtggaga ctgagctcca gggcgtgtgc gacaccgtgc tgggcctgct ggacagccac
     4981 ctcatcaagg aggccgggga cgccgagagc cgggtcttct acctgaagat gaagggtgac
     5041 tactaccgct acctggccga ggtggccacc ggtgacgaca agaagcgcat cattgactca
     5101 gcccggtcag cctaccagga ggccatggac atcagcaaga aggagatgcc gcccaccaac
     5161 cccatccgcc tgggcctggc cctgaacttt tccgtcttcc actacgagat cgccaacagc
     5221 cccgaggagg ccatctctct ggccaagacc actttcgacg aggccatggc tgatctgcac
     5281 accctcagcg aggactccta caaagacagc accctcatca tgcagctgct gcgagacaac
     5341 ctgacactgt ggacggccga caacgccggg gaagaggggg gcgaggctcc ccaggagccc
     5401 cagagctgac tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     5461 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     5521 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     5581 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     5641 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     5701 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     5761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5821 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5881 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5941 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6061 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     6121 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaa
//
