LOCUS       sequence-199001-        5902 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 5902)
  AUTHORS   Gong L, Kamitani T, Fujise K, Caskey LS, Yeh ET
  TITLE     Preferential interaction of sentrin with a ubiquitin-conjugating 
            enzyme, Ubc9.
  JOURNAL   J Biol Chem. 1997 Nov 7;272(45):28198-201.
  PUBMED    9353268
REFERENCE   2  (bases 1 to 5902)
  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."; date: "1997-11-7"; volume: "272(45)"; pages: "28198-201"
COMMENT     Sequence Label: pcDNA3/UBC9
FEATURES             Location/Qualifiers
     source          1..5902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(109..127)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(110..127)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(147..164)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    153..377
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      423..851
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(510..529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     720..741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(860..880)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        865..1194
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1045..1180
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1107..1126
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1261..2055
                     /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(1315..1334)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1925..1944
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2229..2350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2266..2285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2320..2339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2399..2415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2399..2415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2412..2434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2423..2439
                     /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(2447..2453,2454..2471,2472..2477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2492..2513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2630..2647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2801..3389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2881..2900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3560..4351,4352..4420))
                     /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     4183..4202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4421..4525)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4600..4619)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4791..5170
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5171..5374
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5325..5345
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5419..5438
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5419..5437
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaaaagggtc cgagcacaag ccaagaagtt tgcgccctca taagcagcga attctgcaga
       61 tatccatcac actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca
      121 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
      181 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
      241 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
      301 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
      361 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
      421 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      481 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      541 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      601 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
      661 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
      721 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
      781 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
      841 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
      901 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
      961 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1021 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     1081 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     1141 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     1201 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     1261 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     1321 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     1381 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     1441 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     1501 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     1561 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     1621 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     1681 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     1741 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     1801 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     1861 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     1921 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     1981 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     2041 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     2101 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     2161 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     2221 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2281 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2341 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     2401 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2461 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2521 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2581 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2641 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2701 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2761 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2821 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2881 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2941 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3001 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3061 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3121 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3181 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3241 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3301 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3361 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3421 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3481 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3541 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3841 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3901 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3961 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4021 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4081 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4141 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4201 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4261 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4321 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4381 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4441 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4501 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     4561 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     4621 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     4681 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     4741 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
     4801 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     4861 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     4921 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     4981 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     5041 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     5101 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     5161 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     5221 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     5281 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     5341 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
     5401 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc
     5461 tcggatccat gtcggggatc gccctcagca gactcgccca ggagaggaaa gcatggagga
     5521 aagaccaccc atttggtttc gtggctgtcc caacaaaaaa tcccgatggc acgatgaacc
     5581 tcatgaactg ggagtgcgcc attccaggaa agaaagggac tccgtgggaa ggaggcttgt
     5641 ttaaactacg gatgcttttc aaagatgatt atccatcttc gccaccaaaa tgtaaattcg
     5701 aaccaccatt atttcacccg aatgtgtacc cttcggggac agtgtgcctg tccatcttag
     5761 aggaggacaa ggactggagg ccagccatca caatcaaaca gatcctatta ggaatacagg
     5821 aacttctaaa tgaaccaaat atccaagacc cagctcaagc agaggcctac acgatttact
     5881 gccaaaacag agtggagtac ga
//
