LOCUS       sequence-48392-e        8855 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 8855)
  AUTHORS   Nabhan JF, Pan H, Lu Q
  TITLE     Arrestin domain-containing protein 3 recruits the NEDD4 E3 ligase to
            mediate ubiquitination of the beta2-adrenergic receptor.
  JOURNAL   EMBO Rep. 2010 Aug;11(8):605-11. Epub 2010 Jun 18.
  PUBMED    20559325
REFERENCE   2  (bases 1 to 8855)
  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: "EMBO Rep.";
            date: "2010-08"; volume: "11(8)"; pages: "605-11. Epub 2010 Jun 18"
COMMENT     Sequence Label: pCI-neo.mCherry-NEDD4
FEATURES             Location/Qualifiers
     source          1..8855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    45..166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(56..75)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     110..129
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      352..807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(439..458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     649..670
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        924..1281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        924..1115
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(947..967)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1132..1267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1332..2126
                     /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(1386..1405)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1996..2015
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2190..2238
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2279..2298)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2398..2420
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2458..2476)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2544..2648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2649..2717,2718..3509)
                     /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     complement(2867..2886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3680..4268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4169..4188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4479..4858
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4859..5070
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5021..5041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5067..5091
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          5231..5363
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5346..5365
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5408..5426
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5443..6150
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5588..5606)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5856..5875
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 ggccgcttcc ctttagtgag ggttaatgct tcgagcagac atgataagat acattgatga
       61 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      121 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      181 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
      241 cctctacaaa tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa tagcgaagag
      301 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
      361 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
      421 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
      481 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
      541 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
      601 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
      661 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
      721 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
      781 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
      841 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
      901 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
      961 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     1021 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     1081 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     1141 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     1201 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     1261 ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagg
     1321 ctagagccac catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     1381 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     1441 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     1501 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     1561 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     1621 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     1681 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     1741 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1801 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     1861 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1921 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1981 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     2041 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     2101 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     2161 gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt
     2221 gtgttggttt tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc actctcagta
     2281 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     2341 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2401 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     2461 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     2521 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2581 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2641 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2701 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2761 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2821 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2881 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2941 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3001 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3061 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3121 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3181 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3241 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3301 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3361 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3421 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3481 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3541 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3601 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3661 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3721 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3781 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3841 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3901 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3961 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4021 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4081 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4141 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4201 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4261 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4321 ctcacatggc tcgacagatc ttcaatattg gccattagcc atattattca ttggttatat
     4381 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
     4441 catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat tgactagtta
     4501 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     4561 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     4621 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     4681 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc
     4741 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     4801 cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     4861 gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc
     4921 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     4981 tccaaaatgt cgtaacaact gcgatcgccc gccccgttga cgcaaatggg cggtaggcgt
     5041 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cactagaagc
     5101 tttattgcgg tagtttatca cagttaaatt gctaacgcag tcagtgcttc tgacacaaca
     5161 gtctcgaact taagctgcag tgactctctt aaggtagcct tgcagaagtt ggtcgtgagg
     5221 cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
     5281 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
     5341 ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct cttaaggcta
     5401 gagtacttaa tacgactcac tataggctag cctcgagaat tcatggtgag caagggcgag
     5461 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     5521 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     5581 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     5641 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     5701 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     5761 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     5821 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     5881 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     5941 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     6001 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     6061 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     6121 tccaccggcg gcatggacga gctgtacaag atggcaactt gcgcggtgga ggtgttcggg
     6181 ctcctggagg acgaggaaaa ttcacgaatt gtgagagtaa gagttatagc cggaataggc
     6241 cttgccaaga aggatatatt gggagctagt gatccttacg tgagagtgac gttatatgac
     6301 ccaatgaatg gagttcttac aagtgtgcaa acaaaaacca ttaaaaagag tttgaatcca
     6361 aagtggaatg aagaaatatt attcagagtt catcctcagc agcaccggct tctttttgaa
     6421 gtgtttgacg aaaaccgatt gacaagagat gatttcctag gtcaagtgga tgttccactt
     6481 tatccattac cgacagaaaa tccaagattg gagagaccat atacatttaa ggattttgtt
     6541 cttcatccaa gaagtcacaa atcaagagtt aaaggttatc tgagactaaa aatgacttat
     6601 ttacctaaaa ccagtggctc agaagatgat aatgcagaac aggctgagga attagagcct
     6661 ggctgggttg ttttggacca accagatgct gcttgccatt tgcagcaaca acaagaacct
     6721 tctcctctac ctccagggtg ggaagagagg caggatatcc ttggaaggac ctattatgta
     6781 aaccatgaat ctagaagaac acagtggaaa agaccaaccc ctcaggacaa cctaacagat
     6841 gctgagaatg gcaacattca actgcaagca caacgtgcat ttaccaccag gcggcagata
     6901 tccgaggaaa cagaaagtgt tgacaaccga gagtcttccg agaactggga aattataaga
     6961 gaagatgaag ccaccatgta tagcaaccag gccttcccat cacctccacc gtcaagtaac
     7021 ttggatgttc caactcatct tgcagaagaa ttgaatgcca gactcaccat ttttggaaat
     7081 tcagccgtga gccagccagc atcgagctca aatcattcca gcagaagagg cagcttacaa
     7141 gcctatactt ttgaggaaca acctacactt cctgtgcttt tgcctacttc atctggatta
     7201 ccaccaggtt gggaagaaaa acaagatgaa agaggaagat catattatgt agatcacaat
     7261 tccagaacga ctacttggac aaagcccact gtacaggcca cagtggagac cagtcagctg
     7321 acctcaagcc agagttctgc aggccctcaa tcacaagcct ccaccagtga ttcaggccag
     7381 caggtgaccc agccatctga aattgagcaa ggattccttc ctaaaggctg ggaagtccgg
     7441 catgcaccaa atgggaggcc tttctttatt gaccacaaca ctaaaaccac cacctgggaa
     7501 gatccaagat tgaaaattcc agcccatctg agaggaaaga catcacttga tacttccaat
     7561 gatctagggc ctttacctcc aggatgggaa gagagaactc acacagatgg aagaatcttc
     7621 tacataaatc acaatataaa aagaacacaa tgggaagatc ctcggttgga gaatgtagca
     7681 ataactggac cagcagtgcc ctactccagg gattacaaaa gaaagtatga gttcttccga
     7741 agaaagttga agaagcagaa tgacattcca aacaaatttg aaatgaaact tcgccgagca
     7801 actgttcttg aagactctta ccggagaatt atgggtgtca agagagcaga cttcctgaag
     7861 gctcgactgt ggattgagtt tgatggtgaa aagggattgg attatggagg agttgccaga
     7921 gaatggttct tcctgatctc aaaggaaatg tttaaccctt attatgggtt gtttgaatat
     7981 tctgctacgg acaattatac cctacagata aatccaaact ctggattgtg taacgaagat
     8041 cacctctctt acttcaagtt tattggtcgg gtagctggaa tggcagttta tcatggcaaa
     8101 ctgttggatg gttttttcat ccgcccattt tacaagatga tgcttcacaa accaataacc
     8161 cttcatgata tggaatctgt ggatagtgaa tattacaatt ccctaagatg gattcttgaa
     8221 aatgacccaa cagaattgga cctcaggttt atcatagatg aagaactttt tggacagaca
     8281 catcaacatg agctgaaaaa tggtggatca gaaatagttg tcaccaataa gaacaaaaag
     8341 gaatatattt atcttgtaat acaatggcga tttgtaaacc gaatccagaa gcaaatggct
     8401 gcttttaaag agggattctt tgaactaata ccacaggatc tcatcaaaat ttttgatgaa
     8461 aatgaactag agcttcttat gtgtggactg ggagatgttg atgtgaatga ctggagggaa
     8521 catacaaagt ataaaaatgg ctacagtgca aatcatcagg ttatacagtg gttttggaag
     8581 gctgttttaa tgatggattc agaaaaaaga ataagattac ttcagtttgt cactggcaca
     8641 tctcgggtgc ctatgaatgg atttgctgaa ctatacggtt caaatggacc acagtcattt
     8701 acagttgaac agtggggtac tcctgaaaag ctgccaagag ctcatacctg ttttaatcgc
     8761 ctggacttgc caccttatga atcatttgaa gaattatggg ataaacttca gatggcaatt
     8821 gaaaacaccc agggctttga tggagttgat taggc
//
