LOCUS       sequence-270880-        8567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human UBA6 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8567)
  AUTHORS   Pelzer C, Kassner I, Matentzoglu K, Singh RK, Wollscheid HP, 
            Scheffner M, Schmidtke G, Groettrup M
  TITLE     UBE1L2, a novel E1 enzyme specific for ubiquitin.
  JOURNAL   J Biol Chem. 2007 Aug 10;282(32):23010-4. doi: 
            10.1074/jbc.C700111200. Epub 2007 Jun 19.
  PUBMED    17580310
REFERENCE   2  (bases 1 to 8567)
  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"; doi: 
            "10.1074/jbc.C700111200"; journalName: "J Biol Chem"; date: 
            "2007-08-10- 10"; volume: "282"; issue: "32"; pages: "23010-4"
COMMENT     Sequence Label: pcDNA3.1-HA-UBA6
FEATURES             Location/Qualifiers
     source          1..8567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    32..53
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(68..73,74..91,92..98)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    106..122
                     /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)."
     primer_bind     111..133
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     130..146
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     130..146
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    195..316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(206..225)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     260..279
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(490..1284)
                     /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(601..620)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1211..1230
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1351..1680)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1365..1500
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1419..1438)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1665..1685
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1694..2122)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1804..1825)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2016..2035
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2168..2392
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2381..2398
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5604..5630)
                     /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             complement(5604..5630)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5674..5693)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5675..5693)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5738..5941)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5767..5787)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5942..6321
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6493..6512
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6587..6691
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6692..6760,6761..7552)
                     /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     complement(6910..6929)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7723..8311
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8212..8231
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8465..8482
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
       61 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      121 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
      181 gtatacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
      241 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
      301 ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga
      361 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
      421 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
      481 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
      541 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
      601 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
      661 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
      721 atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc
      781 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
      841 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
      901 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
      961 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     1021 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     1081 cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac
     1141 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     1201 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     1261 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatcc
     1321 gaaaatggat atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc
     1381 tcctcactac ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa
     1441 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     1501 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     1561 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     1621 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     1681 attccacaga attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1741 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1801 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1861 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1921 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1981 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2041 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2101 aatgcgccgc tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc
     2161 tcagaagcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc
     2221 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     2281 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     2341 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
     2401 gatcagcggg tttaaacggg ccctctagac tcgagttaat cagtgtcatg actgaagtag
     2461 tatcttactg gaggtcccgg caaatcttca tctccatcaa tgtctggagc aaatgacaca
     2521 gtaagatcca catatttctt ttcagtagta ggttttacaa gtttatgcat tgttaacttc
     2581 aatctttttg catgaccagg cattacagga acataaagca ttttgactcc ctgtaccacc
     2641 attgttggct caattccata cttctctttg actgcattta tgaaatccaa gagggtgaaa
     2701 tcttcttttc catgtacggt ccatcgatcc caaattgtaa atgatattcc atttctgatt
     2761 ttagttttcc ttacttcagt tgtctctgta aatactacaa ttggaatggc taagttaaga
     2821 aaacaatttt tgtaagcttc aaatggatag ccaccagtta ctttgatcat ctccaaggca
     2881 accaagccag aaactgtagc agtggttgtt gctatagcag gtataatttt accagctatg
     2941 cgctttgttt tgaaacggtc agctggttca atgctgtaca ttttggcacg aagatttgat
     3001 gcagctgtga tgaaatctat gtgtccatta tgatcatcat ctttttcaaa tgaaagcact
     3061 gccatctgaa ggtcactttt ggtggcttca ttagataaaa tagccttttc tagttggaaa
     3121 attgcattcc tctcatcttc actgctaata ggaacatggt ctggtttcct tgcagtttca
     3181 tctgtttgaa caaccttatt ggaaggcttg aattcctgaa tctttacttc tgaaagaata
     3241 ttcaagaggg catctgctga taagtcctct tctgcaaatg gaatacaata tactgtagca
     3301 tatagttttg cagcattctg aaggaaactg aggtgcaaag gctcatttaa atcaaatttt
     3361 attggagagg gtggcctctt tggtgactgc caaaataaac tgccatcttt taatcgtatg
     3421 tccagaggga aacagtgaag aagctgaaga gccttatggt taaaatattt ttcaaacttt
     3481 aatcttgcta attctacaca ctgggaccaa tttctaggtc ttctgctaag taactttata
     3541 acttgaaaac agccttctaa actgtgtcca ctctgtatct tctgtaagac ttcttctgca
     3601 gatgaatagg tttgccaaaa tttgttaaac aatgaaggtt tgtgggaaaa ggaactttca
     3661 aacttatctc ttgcccactg tatggtatgt tcaatagcag ctggaaagga ttttagagta
     3721 caaaatggta tttcctcttc tgggggatcc cgatgactat tgtaagactc agtcaaatgc
     3781 ggtacaataa cttcagtgtg tcccttagtg cccattgttc cagaatctaa aagaggcctt
     3841 agatttgcta agcaacgact gtctacgtat ctcctggctt ccacattatc taatgctgta
     3901 ataattacat cttgtttagt atagaactca tcattgtaaa tggtctcagt ggttggacat
     3961 actttgttca ggtgtgcatc tatctttatt tgagaattta ttttcagagt agcatcagca
     4021 gcagtgtagc ttttaggttt ctgtatgtga tgaggacgaa ataggaactg tctatttaag
     4081 ttggatttct ctatcaagtc aggatctgta actgtaatca ttcctttctc tttgcttgtg
     4141 ccaacaccaa gtaaagcaaa atttttcaac atttcacagc ctatggctcc acaccctact
     4201 aagaagatgt ttaaattttg cagtttctga cacaaagtgt ctccaatgca agctcttaag
     4261 gcatcatatc tatctcctcg tgggagaaat tcttcacatt caggtttgcc tagtgattca
     4321 acaatatctg ctgcttcaag atataaccac tggcacaaag gagaaaattt tcctgttaca
     4381 gctttcaata cttcttggct ggcaacacct cctactgctg cagcaagtgg agataaaaag
     4441 ccttgggcag tccaagagag ccaatgcaca atgtcagcat ttacatcagg cttctcttcc
     4501 aaggtttcac ttatagatgt tgctagtttc aacagttctt ctgaatcttg ttggcatcca
     4561 acatttggct tgcgactgta tttctcctga aactggtcca aggcaagcat agctgtgtga
     4621 atctctaaag gtgcctcagg gttgctaaaa tccacaataa ggcactttgg atgttttaac
     4681 tgcctctcca gtgattcaaa aaaaactgtt ttaggagtct taacttggac agctatgcct
     4741 ccatgtaaat atggttccag ttctgtggtg tcaccaatac taaaagaaaa tggcgatatc
     4801 accgttattt gttgtataga tccatttaaa cctgtcattc cattaatttc tcgaaatgtt
     4861 aggaattgtc ctgtctccag tttgtgagga tgattttcaa ggcaagtaac aatgccagga
     4921 tttgcttgcg ttatgtttga aatgaaaatt tcttttggtt cttctcctgt tgtatctaaa
     4981 acttcaaatt catcaccgaa atcacaaaat aaccttgacc aaattccatg tacatctgca
     5041 ctgataaact taattggagg gcactgagaa cggcaaaagt cattgatctt cttctgcaat
     5101 ggaagtttca tctcagtcaa tactacacac tggtatttat ctaaaaagga gagatctgtg
     5161 gtctcattga aaggaacaga agatgatgtg acatgaacgt atggatttag ttctgcaata
     5221 tgtttaagta cagcttcagc cctgtttctc ttattaacaa catcatcttc actgagaaag
     5281 aagttggttc ctagatccca tgcttggcat ttttctgtat catgaattgt aactgcctta
     5341 atccctgcaa gaacaagatt ctttgcaatt tccaaaccaa gaccacccat cccacttaag
     5401 aaaacatggg acttggccat cttctgcatt gctgtgtctc caagaacgta cctctgtcga
     5461 ctatacaatg catcatcgat ttccacagat gctgttgaca taatgggcaa atttttattt
     5521 gtgctgccag tcccccagga agaacaggac gcctcttccc cctgatgggc ggccacaggc
     5581 tcggatcctt ccataggtac cgcggcgtag tcgggcacgt cgtaggggta ggccatggta
     5641 agcttaagtt taaacgctag ccagcttggg tctccctata gtgagtcgta ttaatttcga
     5701 taagccagta agcagtgggt tctctagtta gccagagagc tctgcttata tagacctccc
     5761 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     5821 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     5881 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     5941 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     6001 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     6061 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     6121 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     6181 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     6241 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     6301 aactagtcaa taatcaatgt caacgcgtat atctggcccg tacatcgcga agcagcgcaa
     6361 aacgcctaac cctaagcaga ttcttcatgc aattgtcggt caagccttgc cttgttgtag
     6421 cttaaatttt gctcgcgcac tactcagcga cctccaacac acaagcaggg agcagatact
     6481 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagg ggatcgggag
     6541 atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6601 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6661 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6721 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6781 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6841 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6901 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6961 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     7021 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     7081 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     7141 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7201 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7261 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7321 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7381 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7441 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7501 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7561 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7621 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7681 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7741 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7801 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7861 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7921 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7981 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     8041 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     8101 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     8161 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     8221 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     8281 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     8341 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     8401 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     8461 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     8521 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccg
//
