LOCUS       sequence-364544-        5672 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 5672)
  AUTHORS   Kamitani T, Kito K, Nguyen HP, Yeh ET
  TITLE     Characterization of NEDD8, a developmentally down-regulated 
            ubiquitin-like protein.
  JOURNAL   J Biol Chem. 1997 Nov 7. 272(45):28557-62.
  PUBMED    9353319
REFERENCE   2  (bases 1 to 5672)
  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. 1997 Nov 7. 272(45):28557-62."
COMMENT     Sequence Label: HA-NEDD8
FEATURES             Location/Qualifiers
     source          1..5672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(82..102)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        87..416
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      267..402
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     329..348
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             483..1277
                     /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(537..556)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1147..1166
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1451..1572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1488..1507)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1542..1561
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1621..1637)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1621..1637)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1634..1656)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1645..1661
                     /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(1669..1675,1676..1693,1694..1699))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1714..1735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1852..1869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2023..2611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2103..2122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2782..3573,3574..3642))
                     /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     3405..3424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3643..3747)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3822..3841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4013..4392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4393..4596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4547..4567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4641..4660
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4641..4659
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4676..4702
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(5003..5021)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5004..5021)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5041..5058)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5047..5271
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5317..5672,1..73)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5404..5423)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5614..5635
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
       61 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
      121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
      181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      241 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
      301 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
      361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
      421 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
      481 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
      541 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
      601 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
      661 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
      721 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
      781 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
      841 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
      901 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
      961 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     1021 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     1081 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     1141 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     1201 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     1261 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     1321 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     1381 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     1441 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1501 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1561 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     1621 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1681 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1741 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1861 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1921 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2461 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2701 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2761 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2821 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2881 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2941 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3001 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3061 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3121 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3181 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3241 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3301 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3361 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3421 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3481 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3541 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3601 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3661 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3721 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     3781 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     3841 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     3901 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     3961 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     4021 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     4081 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     4141 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     4201 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     4261 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     4321 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     4381 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     4441 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     4501 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     4561 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     4621 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttatgtaccc
     4681 atacgatgtt ccggattacg ctagcggatc catgctaatt aaagtgaaga cgctgaccgg
     4741 aaaggagatt gagattgaca ttgaacctac agacaaggtg gagcgaatca aggagcgtgt
     4801 ggaggagaaa gagggaatcc ccccacaaca gcagaggctc atctacagtg gcaagcagat
     4861 gaatgatgag aagacagcag ctgattacaa gattttaggt ggttcagtcc ttcacctggt
     4921 gttggctctg agaggaggat gagaattctg cagatatcca tcacactggc ggccgctcga
     4981 gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag
     5041 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     5101 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     5161 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     5221 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     5281 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     5341 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5401 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5461 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5521 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5581 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5641 cactcaaccc tatctcggtc tattcttttg at
//
