LOCUS       sequence-197571-        5377 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 5377)
  AUTHORS   Yasugi T, Howley PM
  TITLE     Identification of the structural and functional human homolog of the
            yeast ubiquitin conjugating enzyme UBC9.
  JOURNAL   Nucleic Acids Res. 1996 Jun 1. 24(11):2005-10.
  PUBMED    8668529
REFERENCE   2  (bases 1 to 5377)
  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: "Nucleic 
            Acids Res. 1996 Jun 1. 24(11):2005-10."
COMMENT     Sequence Label: p3259 pCMV hUBC9 mt HA
FEATURES             Location/Qualifiers
     source          1..5377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        178..557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     712..732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1317..1343
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1405..2027
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1604..1623)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2051..2071)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2056..2385
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2236..2371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2424..2520
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(2596..2615)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2597..2615)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2629..2645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2629..2645)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2642..2664)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2653..2669
                     /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(2677..2683,2684..2701,2702..2707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2722..2743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2860..2877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3031..3619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3111..3130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3790..4581,4582..4650))
                     /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     4413..4432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4651..4755)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4782..5237
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4869..4888)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5079..5100
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(5363..5377,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5377,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
       61 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
      121 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
      181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      421 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      481 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
      541 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
      601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      661 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
      721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      781 attgttttta tttttaattt tctttcaaat acttccatcg aattcagatc ttgaacatgt
      841 cggggatcgc cctcagcaga ctcgcccagg agaggaaagc atggaggaaa gaccacccat
      901 ttggtttcgt ggctgtccca acaaaaaatc ccgatggcac gatgaacctc atgaactggg
      961 agtgcgccat tccaggaaag aaagggactc cgtgggaagg aggcttgttt aaactacgga
     1021 tgcttttcaa agatgattat ccatcttcgc caccaaaatg taaattcgaa ccaccattat
     1081 ttcacccgaa tgtgtaccct tcggggacag tgagcctgtc catcttagag gaggacaagg
     1141 actggaggcc agccattaca atcaagcaga tcctattagg aatacaggaa cttctaaatg
     1201 aaccaaatat ccaagaccca gctcaagcag aggcctacac gatttactgc caaaacagag
     1261 tggagtacga gaaaagggtc cgagcacaag ccaagaagtt tgcgcccgca ccgcggtacc
     1321 cttatgatgt gccagattat gcctgatctg gtaccacgcg tatcgataag cttgcatgcc
     1381 tgcaggtcga ctctagagga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     1441 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     1501 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     1561 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     1621 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     1681 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     1741 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     1801 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     1861 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     1921 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     1981 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     2041 gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2101 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2161 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2221 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2281 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2341 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa
     2401 ctgaaaaacc agaaagttaa ctggtaagtt tagtcttttt gtcttttatt tcaggtcccg
     2461 gatccggtgg tggtgcaaat caaagaactg ctcctcagtg gatgttgcct ttacttctag
     2521 gcctgtacgg aagtgttact tctgctctaa aagctgctgc aggtcgactc tagggccgcg
     2581 taccgagctc gaattcccta tagtgagtcg tattaaattc gtaatcatgt catagctgtt
     2641 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2701 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2761 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2821 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2881 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2941 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3001 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3061 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3121 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3181 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3241 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3301 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3361 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3421 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3481 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3541 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3601 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3661 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3721 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3781 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3841 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3901 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3961 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4021 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4081 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4141 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4201 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4261 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4321 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4381 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4441 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4501 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4561 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4621 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4681 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4741 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     4801 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4861 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4921 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4981 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5041 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5101 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5161 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5221 attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     5281 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     5341 agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
