LOCUS       sequence-365386-        5885 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 5885)
  AUTHORS   Kim KI, Giannakopoulos NV, Virgin HW, Zhang DE
  TITLE     Interferon-inducible ubiquitin E2, Ubc8, is a conjugating enzyme for
            protein ISGylation.
  JOURNAL   Mol Cell Biol. 2004 Nov . 24(21):9592-600.
  PUBMED    15485925
REFERENCE   2  (bases 1 to 5885)
  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: "Mol Cell 
            Biol. 2004 Nov . 24(21):9592-600."
COMMENT     Sequence Label: pcDNA-hISG15
FEATURES             Location/Qualifiers
     source          1..5885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(139..159)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        144..473
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      324..459
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     386..405
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             540..1334
                     /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(594..613)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1204..1223
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1508..1629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1545..1564)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1599..1618
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1678..1694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1678..1694)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1691..1713)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1702..1718
                     /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(1726..1732,1733..1750,1751..1756))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1771..1792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1909..1926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2080..2668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2160..2179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2839..3630,3631..3699))
                     /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     3462..3481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3700..3804)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3879..3898)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4070..4449
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4450..4653
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4604..4624
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4698..4717
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4698..4716
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5269..5285)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5269..5285)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5311..5328)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5317..5541
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5587..5885,1..130)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5674..5693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(5884..5885,1..20)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
       61 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      121 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      181 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      241 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      301 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      361 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
      421 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
      481 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
      541 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
      601 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
      661 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
      721 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
      781 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
      841 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
      901 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
      961 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     1021 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     1081 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     1141 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     1201 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     1261 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     1321 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     1381 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     1441 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     1501 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1561 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1621 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     1681 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1741 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1801 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1861 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2521 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2881 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2941 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3001 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3061 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3121 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3181 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3241 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3301 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3361 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3421 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     3841 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     3901 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     3961 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     4021 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     4081 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     4141 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     4201 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     4261 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     4321 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     4381 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     4441 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     4501 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4561 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4621 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
     4681 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
     4741 aacttaagct tgatggaatt ccatatgggc tgggacctga cggtgaagat gctggcgggc
     4801 aacgaattcc aggtgtccct gagcagctcc atgtcggtgt cagagctgaa ggcgcagatc
     4861 acccagaaga tcggcgtgca cgccttccag cagcgtctgg ctgtccaccc gagcggtgtg
     4921 gcgctgcagg acagggtccc ccttgccagc cagggcctgg gccccggcag cacggtcctg
     4981 ctggtggtgg acaaatgcga cgaacctctg agcatcctgg tgaggaataa caagggccgc
     5041 agcagcacct acgaggtgcg gctgacgcag accgtggccc acctgaagca gcaagtgagc
     5101 gggctggagg gtgtgcagga cgacctgttc tggctgacct tcgaggggaa gcccctggag
     5161 gaccagctcc cgctggggga gtacggcctc aagcccctga gcaccgtgtt catgaatctg
     5221 cgcctgaggg gtgggtaagg atccgaatcg aattcctgca gcccggggga tccactagtt
     5281 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     5341 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     5401 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     5461 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     5521 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     5581 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5641 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5701 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5761 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5821 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5881 atagt
//
