LOCUS       sequence-359427-        7074 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 7074)
  AUTHORS   Hsu SC, Hung MC
  TITLE     Characterization of a novel tripartite nuclear localization sequence
            in the EGFR family.
  JOURNAL   J Biol Chem. 2007 Apr 6;282(14):10432-40. Epub 2007 Feb 5.
  PUBMED    17283074
REFERENCE   2  (bases 1 to 7074)
  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."; date: "2007-04-6"; volume: "282(14)"; pages: "10432-40. Epub
            2007 Feb 5"
COMMENT     Sequence Label: pcDNA6A-EGFR ECD (1-644)
FEATURES             Location/Qualifiers
     source          1..7074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2945..2974
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2990..3007
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3030..3047)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3036..3260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3306..3734
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3393..3412)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3603..3624
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3743..3763)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3748..4077
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3928..4063
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3990..4009
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4125..4172
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4191..4589
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4747..4868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4784..4803)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4838..4857
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4917..4933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4917..4933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4930..4952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4941..4957
                     /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(4965..4971,4972..4989,4990..4995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5010..5031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5148..5165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5319..5907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5399..5418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6078..6869,6870..6938))
                     /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     6701..6720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6939..7043)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttcgg ggagcagcga tgcgaccctc cgggacggcc ggggcagcgc tcctggcgct
      961 gctggctgcg ctctgcccgg cgagtcgggc tctggaggaa aagaaagttt gccaaggcac
     1021 gagtaacaag ctcacgcagt tgggcacttt tgaagatcat tttctcagcc tccagaggat
     1081 gttcaataac tgtgaggtgg tccttgggaa tttggaaatt acctatgtgc agaggaatta
     1141 tgatctttcc ttcttaaaga ccatccagga ggtggctggt tatgtcctca ttgccctcaa
     1201 cacagtggag cgaattcctt tggaaaacct gcagatcatc agaggaaata tgtactacga
     1261 aaattcctat gccttagcag tcttatctaa ctatgatgca aataaaaccg gactgaagga
     1321 gctgcccatg agaaatttac aggaaatcct gcatggcgcc gtgcggttca gcaacaaccc
     1381 tgccctgtgc aacgtggaga gcatccagtg gcgggacata gtcagcagtg actttctcag
     1441 caacatgtcg atggacttcc agaaccacct gggcagctgc caaaagtgtg atccaagctg
     1501 tcccaatggg agctgctggg gtgcaggaga ggagaactgc cagaaactga ccaaaatcat
     1561 ctgtgcccag cagtgctccg ggcgctgccg tggcaagtcc cccagtgact gctgccacaa
     1621 ccagtgtgct gcaggctgca caggcccccg ggagagcgac tgcctggtct gccgcaaatt
     1681 ccgagacgaa gccacgtgca aggacacctg ccccccactc atgctctaca accccaccac
     1741 gtaccagatg gatgtgaacc ccgagggcaa atacagcttt ggtgccacct gcgtgaagaa
     1801 gtgtccccgt aattatgtgg tgacagatca cggctcgtgc gtccgagcct gtggggccga
     1861 cagctatgag atggaggaag acggcgtccg caagtgtaag aagtgcgaag ggccttgccg
     1921 caaagtgtgt aacggaatag gtattggtga atttaaagac tcactctcca taaatgctac
     1981 gaatattaaa cacttcaaaa actgcacctc catcagtggc gatctccaca tcctgccggt
     2041 ggcatttagg ggtgactcct tcacacatac tcctcctctg gatccacagg aactggatat
     2101 tctgaaaacc gtaaaggaaa tcacagggtt tttgctgatt caggcttggc ctgaaaacag
     2161 gacggacctc catgcctttg agaacctaga aatcatacgc ggcaggacca agcaacatgg
     2221 tcagttttct cttgcagtcg tcagcctgaa cataacatcc ttgggattac gctccctcaa
     2281 ggagataagt gatggagatg tgataatttc aggaaacaaa aatttgtgct atgcaaatac
     2341 aataaactgg aaaaaactgt ttgggacctc cggtcagaaa accaaaatta taagcaacag
     2401 aggtgaaaac agctgcaagg ccacaggcca ggtctgccat gccttgtgct cccccgaggg
     2461 ctgctggggc ccggagccca gggactgcgt ctcttgccgg aatgtcagcc gaggcaggga
     2521 atgcgtggac aagtgcaacc ttctggaggg tgagccaagg gagtttgtgg agaactctga
     2581 gtgcatacag tgccacccag agtgcctgcc tcaggccatg aacatcacct gcacaggacg
     2641 gggaccagac aactgtatcc agtgtgccca ctacattgac ggcccccact gcgtcaagac
     2701 ctgcccggca ggagtcatgg gagaaaacaa caccctggtc tggaagtacg cagacgccgg
     2761 ccatgtgtgc cacctgtgcc atccaaactg cacctacgga tgcactgggc caggtcttga
     2821 aggctgtcca acgaatgggc ctaagatccc gtccatcgcc actgggatgg tgggggccct
     2881 cctcttgctg ctggtggtgg ccctggggat cggcctcttc atgctcgagt ctagagggcc
     2941 cttcgaacaa aaactcatct cagaagagga tctgaatatg cataccggtc atcatcacca
     3001 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3061 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3121 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3181 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3241 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3301 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3361 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3421 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3481 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3541 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3601 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3661 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3721 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3781 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3841 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3901 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3961 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4021 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4081 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
     4141 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagc
     4201 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc
     4261 ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca
     4321 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca
     4381 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca
     4441 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga
     4501 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc
     4561 tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca ggactgacac
     4621 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4681 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4741 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4801 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4861 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4921 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4981 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5041 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5101 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5161 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5221 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5281 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5341 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5401 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5461 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5521 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5581 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5641 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5701 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5761 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5821 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5881 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5941 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6001 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6061 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6121 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6181 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6241 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6301 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6361 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6421 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6481 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6541 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6601 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6661 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6721 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6781 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6841 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6901 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6961 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7021 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
