LOCUS       sequence-363016-        7419 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 7419)
  TITLE     SHP2 plasmids
REFERENCE   2  (bases 1 to 7419)
  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"
COMMENT     Sequence Label: pCMV-SHP2 WT
FEATURES             Location/Qualifiers
     source          1..7419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(131..151)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        136..465
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      316..451
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     378..397
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             532..1326
                     /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(586..605)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1196..1215
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1500..1621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1537..1556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1591..1610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1670..1686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1670..1686)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1683..1705)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1694..1710
                     /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(1718..1724,1725..1742,1743..1748))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1763..1784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1901..1918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2072..2660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2152..2171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2831..3622,3623..3691))
                     /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     3454..3473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3692..3796)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3871..3890)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4062..4441
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4442..4645
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4596..4616
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4690..4709
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4690..4708
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6799..6817)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6800..6817)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6837..6854)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6843..7067
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7113..7419,1..122)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7200..7219)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(7410..7419,1..12)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
       61 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      121 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
      181 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
      241 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
      301 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
      361 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
      421 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
      481 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
      541 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
      601 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
      661 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
      721 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
      781 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
      841 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
      901 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
      961 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     1021 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     1081 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     1141 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     1201 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     1261 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     1321 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     1381 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     1441 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     1501 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1561 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1621 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     1681 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1741 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1801 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1861 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1921 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1981 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2041 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2101 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2161 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2221 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2281 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2341 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2401 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2461 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2521 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2581 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2641 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2701 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2761 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2821 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2881 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2941 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3001 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3061 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3121 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3181 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3241 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3301 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3361 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3421 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3481 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3541 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3601 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3661 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3721 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3781 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     3841 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3901 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     3961 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     4021 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     4081 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     4141 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     4201 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4261 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4321 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     4381 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     4441 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     4501 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     4561 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     4621 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
     4681 tatcgaaatt aatacgactc actataggga gacccaagct tggtaccgag ctcggatcca
     4741 ctagtaacgg ccgccagtgt gctggaattc cgggcgggag gaacatgaca tcgcggagat
     4801 ggtttcaccc aaatatcact ggtgtggagg cagaaaacct actgttgaca agaggagttg
     4861 atggcagttt tttggcaagg cctagtaaaa gtaaccctgg agacttcaca ctttccgtta
     4921 gaagaaatgg agctgtcacc cacatcaaga ttcagaacac tggtgattac tatgacctgt
     4981 atggagggga gaaatttgcc actttggctg agttggtcca gtattacatg gaacatcacg
     5041 ggcaattaaa agagaagaat ggagatgtca ttgagcttaa atatcctctg aactgtgcag
     5101 atcctacctc tgaaaggtgg tttcatggac atctctctgg gaaagaagca gagaaattat
     5161 taactgaaaa aggaaaacat ggtagttttc ttgtacgaga gagccagagc caccctggag
     5221 attttgttct ttctgtgcgc actggtgatg acaaagggga gagcaatgac ggcaagtcta
     5281 aagtgaccca tgttatgatt cgctgtcagg aactgaaata cgacgttggt ggaggagaac
     5341 ggtttgattc tttgacagat cttgtggaac attataagaa gaatcctatg gtggaaacat
     5401 tgggtacagt actacaactc aagcagcccc ttaacacgac tcgtataaat gctgctgaaa
     5461 tagaaagcag agttcgagaa ctaagcaaat tagctgagac cacagataaa gtcaaacaag
     5521 gcttttggga agaatttgag acactacaac aacaggagtg caaacttctc tacagccgaa
     5581 aagagggtca aaggcaagaa aacaaaaaca aaaatagata taaaaacatc ctgccctttg
     5641 atcataccag ggttgtccta cacgatggtg atcccaatga gcctgtttca gattacatca
     5701 atgcaaatat catcatgcct gaatttgaaa ccaagtgcaa caattcaaag cccaaaaaga
     5761 gttacattgc cacacaaggc tgcctgcaaa acacggtgaa tgacttttgg cggatggtgt
     5821 tccaagaaaa ctcccgagtg attgtcatga caacgaaaga agtggagaga ggaaagagta
     5881 aatgtgtcaa atactggcct gatgagtatg ctctaaaaga atatggcgtc atgcgtgtta
     5941 ggaacgtcaa agaaagcgcc gctcatgact atacgctaag agaacttaaa ctttcaaagg
     6001 ttggacaagg gaatacggag agaacggtct ggcaatacca ctttcggacc tggccggacc
     6061 acggcgtgcc cagcgaccct gggggcgtgc tggacttcct ggaggaggtg caccataagc
     6121 aggagagcat catggatgca gggccggtcg tggtgcactg cagtgctgga attggccgga
     6181 cagggacgtt cattgtgatt gatattctta ttgacatcat cagagagaaa ggtgttgact
     6241 gcgatattga cgttcccaaa accatccaga tggtgcggtc tcagaggtca gggatggtcc
     6301 agacagaagc acagtaccga tttatctata tggcggtcca gcattatatt gaaacactac
     6361 agcgcaggat tgaagaagag cagaaaagca agaggaaagg gcacgaatat acaaatatta
     6421 agtattctct agcggaccag acgagtggag atcagagccc tctcccgcct tgtactccaa
     6481 cgccaccctg tgcagaaatg agagaagaca gtgctagagt ctatgaaaac gtgggcctga
     6541 tgcaacagca gaaaagtttc agatgagaaa acctgccaaa acttcagcac agaaatagat
     6601 gtggactttc accctctccc taaaaagatc aagaacagac gcaagaaagt ttatgtgaag
     6661 acagaatttg gatttggaag gcttgcaatg tggttgacta ccttttgata agcaaaattt
     6721 gaaaccattt aaagaccgga attctgcaga tatccatcac actggcggcc gctcgagcat
     6781 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     6841 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     6901 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     6961 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     7021 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     7081 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     7141 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     7201 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     7261 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     7321 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     7381 tttgacgttg gagtccacgt tctttaatag tggactctt
//
