LOCUS       sequence-190048-        7956 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 7956)
  AUTHORS   Sun Y, Liu X, Ng-Eaton E, Lodish HF, Weinberg RA
  TITLE     SnoN and Ski protooncoproteins are rapidly degraded in response to 
            transforming growth factor beta signaling.
  JOURNAL   Proc Natl Acad Sci U S A. 1999 Oct 26. 96(22):12442-7.
  PUBMED    10535941
REFERENCE   2  (bases 1 to 7956)
  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: "Proc Natl 
            Acad Sci U S A. 1999 Oct 26. 96(22):12442-7."
COMMENT     Sequence Label: pCI-Neo HA-hSki
FEATURES             Location/Qualifiers
     source          1..7956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        188..567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        568..779
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     730..750
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     776..800
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          940..1072
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1055..1074
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1117..1135
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1159..1185
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3710..3831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3721..3740)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3775..3794
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4017..4472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4104..4123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4314..4335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4589..4946
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4589..4780
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4612..4632)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4797..4932
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4997..5791
                     /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(5051..5070)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5661..5680
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5855..5903
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5944..5963)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6063..6085
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6123..6141)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6209..6313
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6314..6382,6383..7174)
                     /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(6532..6551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7345..7933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7834..7853
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct tcaatattgg
       61 ccattagcca tattattcat tggttatata gcataaatca atattggcta ttggccattg
      121 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
      181 ccatgttggc attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      241 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      301 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      361 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      421 gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg
      481 cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc
      541 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt
      601 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      661 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg cgatcgcccg
      721 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      781 gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac agttaaattg
      841 ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt gactctctta
      901 aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga
      961 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     1021 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac
     1081 tcccagttca attacagctc ttaaggctag agtacttaat acgactcact ataggctagc
     1141 ctcgagacca ccatggggta cccatacgat gtcccagact acgcttccat ggaggcggcg
     1201 gcaggcggcc gcggctgttt ccagccgcac ccggggctgc agaagacgct ggagcagttc
     1261 cacctgagct ccatgagctc gctgggcggc ccggccgctt tctcggcgcg ctgggcgcag
     1321 gaggcctaca agaaggagag cgccaaggag gcgggcgcgg ccgcggtgcc ggcgccggtg
     1381 cccgcagcca ccgagccgcc gcccgtgctg cacctgcccg ccatccagcc gccgccgccc
     1441 gtgctgcccg ggcccttctt catgccgtcc gaccgctcca ccgagcgctg cgagaccgta
     1501 ctggaaggcg agaccatctc gtgcttcgtg gtgggaggcg agaagcgcct gtgtctgccg
     1561 cagattctca actcggtgct gcgcgacttc tcgctgcagc agatcaacgc ggtgtgcgac
     1621 gagctccaca tctactgctc gcgctgcacg gccgaccagc tggagatcct caaagtcatg
     1681 ggcatcctgc ccttctcggc gccctcgtgc gggctcatca ccaagacgga cgccgagcgc
     1741 ctgtgcaacg cgctgctcta cggcggcgcc tacccgccgc cctgcaagaa ggagctggcc
     1801 gccagcctgg cgctgggcct ggagctcagc gagcgcagcg tccgcgtgta ccacgagtgc
     1861 ttcggcaagt gtaaggggct gctggtgccc gagctctaca gcagcccgag cgccgcctgc
     1921 atccagtgcc tggactgccg cctcatgtac ccgccgcaca agttcgtggt gcactcgcac
     1981 aaggccctgg agaaccggac ctgccactgg ggcttcgact cggccaactg gcgggcctac
     2041 atcctgctga gccaggatta cacgggcaag gaggagcagg cgcgcctcgg ccgctgcctg
     2101 gacgacgtga aggagaaatt cgactatggc aacaagtaca agcggcgggt gccccgggtc
     2161 tcctctgagc ctccggcctc cataagaccc aaaacagatg acacctcttc ccagtccccc
     2221 gcgccttccg aaaaggacaa gccgtccagc tggctgcgga ccttggccgg ctcttccaat
     2281 aagagcctgg gctgtgttca ccctcgccag cgcctctctg ctttccgacc ctggtccccc
     2341 gcagtgtcag cgagtgagaa agagctctcc ccacacctcc cggccctcat ccgagacagc
     2401 ttctactcct acaagagctt tgagacagcc gtggcgccca acgtggccct cgcaccgccg
     2461 gcccagcaga aggttgtgag cagccctccg tgtgccgccg ccgtctcccg ggcccccgag
     2521 cctctcgcca cttgcaccca gcctcggaag cggaagctga ctgtggacac cccaggagcc
     2581 ccagagacgc tggcgcccgt ggctgcccca gaggaggaca aggactcgga ggcggaggtg
     2641 gaagttgaaa gcagggagga attcacctcc tccttgtcct cgctctcttc cccgtccttt
     2701 acctcatcca gctccgccaa ggacctgggc tccccgggtg cgcgtgccct gccctcggcc
     2761 gtccctgatg ctgcggcccc tgccgacgcc cccagtgggc tggaggcgga gctggagcac
     2821 ctgcggcagg cactggaggg cggcctggac accaaggaag ccaaagagaa gttcctgcat
     2881 gaggtggtca agatgcgcgt gaagcaggag gagaagctca gcgcagccct gcaggccaag
     2941 cgcagcctcc accaggagct ggagttccta cgcgtggcca agaaggagaa gctgcgggag
     3001 gccacggagg ccaagcgtaa cctgcggaag gagatcgagc gtctccgcgc cgagaacgag
     3061 aagaagatga aagaggccaa cgagtcacgg ctgcgcctga agcgggagct ggagcaggcg
     3121 cggcaggccc gggtgtgcga caagggctgc gaggcgggcc gcctgcgcgc caagtactcg
     3181 gcccagatcg aagacctgca ggtgaagctg cagcacgcgg aggcggaccg ggagcagctg
     3241 cgggccgacc tgctgcggga gcgcgaggcc cgggagcacc tggagaaggt ggtgaaggag
     3301 ctgcaggaac agctgtggcc gcgggcccgc cccgaggctg cgggcagcga gggcgctgcg
     3361 gagctggagc cgtagattcc gtgcctgccg ccgcagcgcc gccgacaacg cgggtgcagg
     3421 ggggcgcggc tgggcggtgc agctccgccc ggctccgccc ctgcagccca cacagcacaa
     3481 cgtcttaccg tgcctattac caagcgagtg tttgtaacca tgtagttttg gaacccactg
     3541 caaaattttc tactggccaa gttcaagtga gtaagccgcg tcccccaact acagctggag
     3601 acggggccag ctcggcggcc tgctggtcct ctgcttgctg gaacaggtag cagtccggat
     3661 cgggcggccg cttcccttta gtgagggtta atgcttcgag cagacatgat aagatacatt
     3721 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3781 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3841 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     3901 taaaacctct acaaatgtgg taaaatccga taaggatcga tccgggctgg cgtaatagcg
     3961 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc
     4021 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4081 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4141 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4201 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4261 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4321 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4381 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4441 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     4501 ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct
     4561 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
     4621 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4681 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4741 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4801 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4861 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4921 tttttggagg cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact
     4981 taaggctaga gccaccatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5041 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5101 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5161 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5221 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5281 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5341 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5401 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5461 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5521 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5581 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5641 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5701 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5761 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5821 caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     5881 tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg atccgcgtat ggtgcactct
     5941 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     6001 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     6061 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     6121 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     6181 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6241 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6301 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6361 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6421 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6481 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6541 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6601 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6661 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6721 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6781 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6841 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6901 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6961 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7021 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7081 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7141 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7201 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7261 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7321 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7381 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7441 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7501 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7561 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7621 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7681 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7741 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7801 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7861 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7921 gagcctatgg aaaaacgcca gcaacgcggc cttttt
//
