LOCUS       sequence-166216-        5378 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 5378)
  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 5378)
  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: p3258 pCMV hUBC9 wt HA
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1933..2388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2020..2039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2230..2251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2514..2536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2528..2545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2706..3085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3086..3289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3240..3260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3846..3872
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3934..4556
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4133..4152)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4580..4600)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4585..4914
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4765..4900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4953..5049
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(5125..5144)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5126..5144)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5158..5174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5158..5174)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5171..5193)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5182..5198
                     /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(5206..5212,5213..5230,5231..5236))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5251..5272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1981 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2221 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2281 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2341 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     2581 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     2641 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     2701 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     2761 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2821 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2881 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2941 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     3001 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     3061 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     3121 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3181 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     3241 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     3301 ccgtcagaat tgtttttatt tttaattttc tttcaaatac ttccatcgaa ttcagatctt
     3361 tgaacatgtc ggggatcgcc ctcagcagac tcgcccagga gaggaaagca tggaggaaag
     3421 accacccatt tggtttcgtg gctgtcccaa caaaaaatcc cgatggcacg atgaacctca
     3481 tgaactggga gtgcgccatt ccaggaaaga aagggactcc gtgggaagga ggcttgttta
     3541 aactacggat gcttttcaaa gatgattatc catcttcgcc accaaaatgt aaattcgaac
     3601 caccattatt tcacccgaat gtgtaccctt cggggacagt gtgcctgtcc atcttagagg
     3661 aggacaagga ctggaggcca gccatcacaa tcaaacagat cctattagga atacaggaac
     3721 ttctaaatga accaaatatc caagacccag ctcaagcaga ggcctacacg atttactgcc
     3781 aaaacagagt ggagtacgag aaaagggtcc gagcacaagc caagaagttt gcgccctcac
     3841 cgcggtaccc ttatgatgtg ccagattatg cctgatctgg taccacgcgt atcgataagc
     3901 ttgcatgcct gcaggtcgac tctagaggat cccgggtggc atccctgtga cccctcccca
     3961 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4021 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4081 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4141 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     4201 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     4261 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     4321 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     4381 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag
     4441 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc
     4501 ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt
     4561 acgcggccag cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4621 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4681 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4741 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4801 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4861 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4921 ctcgaggaac tgaaaaacca gaaagttaac tggtaagttt agtctttttg tcttttattt
     4981 caggtcccgg atccggtggt ggtgcaaatc aaagaactgc tcctcagtgg atgttgcctt
     5041 tacttctagg cctgtacgga agtgttactt ctgctctaaa agctgctgca ggtcgactct
     5101 agggccgcgt accgagctcg aattccctat agtgagtcgt attaaattcg taatcatgtc
     5161 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5221 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5281 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5341 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgct
//
