LOCUS       sequence-251048-        5138 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 5138)
  AUTHORS   Zou W, Papov V, Malakhova O, Kim KI, Dao C, Li J, Zhang DE
  TITLE     ISG15 modification of ubiquitin E2 Ubc13 disrupts its ability to 
            form thioester bond with ubiquitin.
  JOURNAL   Biochem Biophys Res Commun. 2005 Oct 14. 336(1):61-8.
  PUBMED    16122702
REFERENCE   2  (bases 1 to 5138)
  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: "Biochem 
            Biophys Res Commun. 2005 Oct 14. 336(1):61-8."
COMMENT     Sequence Label: pFlagCMV2-UbcH13
FEATURES             Location/Qualifiers
     source          1..5138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..62)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        47..376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      227..362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(414..433)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(415..433)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(447..463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(447..463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(460..482)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    471..487
                     /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(495..501,502..519,520..525))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    540..561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(678..695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(849..1437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(929..948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1608..2399,2400..2468))
                     /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     2231..2250
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2469..2573)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2600..3055
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2687..2706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2897..2918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3181..3203
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3195..3212
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3196..3212
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3373..3752
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3753..3956
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3907..3927
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3990..4013
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    join(4534..5138,1..18)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4733..4752)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 gtagatgcct gttgaattgg gtacgcggcc agcttggctg tggaatgtgt gtcagttagg
       61 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      121 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
      181 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
      241 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
      301 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
      361 cctaggcttt tgcaaaaagc tcctcgagga actgaaaaac cagaaagtta attccctata
      421 gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
      481 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
      541 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
      601 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
      661 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
      721 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
      781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      841 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      901 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      961 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1021 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1081 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1141 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1201 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1261 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1321 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1381 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1441 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1501 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1561 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1621 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1681 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1741 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1801 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     1861 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1921 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1981 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2041 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2101 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2161 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2221 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2281 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2341 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2401 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     2461 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     2521 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2581 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2641 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2701 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2761 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2821 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2881 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2941 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3001 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttccatt
     3061 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     3121 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     3181 cccagtcacg acgttgtaaa acgacggcca gtgccaagct gatctataca ttgaatcaat
     3241 attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc
     3301 cattgcatac gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat
     3361 gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     3421 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     3481 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     3541 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     3601 tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta
     3661 aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt
     3721 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg
     3781 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     3841 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc
     3901 cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     3961 tagtgaaccg tcagaattga tctaccatgg actacaaaga cgatgacgac aagcttatgg
     4021 ccgggctgcc ccgcaggatc atcaaggaaa cccagcgttt gctggcagaa ccagttcctg
     4081 gcatcaaagc cgaaccagat gagagcaacg cccgttattt tcatgtggtc attgctggcc
     4141 ctcaggattc cccctttgag ggagggactt ttaaacttga actattcctt ccagaagaat
     4201 acccaatggc agcccctaaa gtacgtttca tgaccaaaat ttatcatcct aatgtagaca
     4261 agttgggaag aatatgttta gatattttga aagataagtg gtccccagca ctgcagatcc
     4321 gcacagttct gctatcgatc caggccttgt taagtgctcc caatccagat gatccattag
     4381 caaatgatgt agcggagcag tggaagacca acgaagccca agccatagaa acagctagag
     4441 catggactag gctatatgcc atgaataata tttaagcggc cgcgaattca tcgatagatc
     4501 tgatatcggt accagtcgac tctagaggat cccgggtggc atccctgtga cccctcccca
     4561 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4621 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4681 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4741 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     4801 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     4861 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     4921 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     4981 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag
     5041 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc
     5101 ttgcttggca ctgtcctctc atgcgttggg tccactca
//
