LOCUS       Exported                5466 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCS2-myc-gfp-dSH2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5466)
  AUTHORS   Robles E, Woo S, Gomez TM
  TITLE     Src-dependent tyrosine phosphorylation at the tips of growth cone 
            filopodia promotes extension.
  JOURNAL   J Neurosci. 2005 Aug 17;25(33):7669-81.
  PUBMED    16107653
REFERENCE   2  (bases 1 to 5466)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5466
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             103..132
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             139..855
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             139..855
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(184..205)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(445..464)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     792..813
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1493..1510)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1515..1649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1539..1558)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1593..1612
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1762..1782)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1762..1780)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1801..1817)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1801..1817)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1814..1836)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1825..1841
                     /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(1849..1879)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1894..1915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2032..2049)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2203..2791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2283..2302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2962..3822)
                     /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     3585..3604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3823..3927)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3953..4408)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4090..4111)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4302..4321
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4482..5466
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tttaaagcta tggagcaaaa gctcatttct
      121 gaagaggact tgaatgaaat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      181 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      241 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      301 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      361 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      421 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      481 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      541 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      601 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      661 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      721 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
      781 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      841 gacgagctgt acaaggaatt tcgagatcca ggctgaggag tggtactttg gcaagatcac
      901 tagacgggaa tcagagcggc tgctgctcaa cgccgagaac ccgagaggga ccttcctcgt
      961 gagggagagt gagaccacaa aaggtgccta ctgcctctct gtatccgact tcgacaatgc
     1021 caagggccta aatgtgaaac actacaagat ccgcaagctg gacagcggcg gtttctacat
     1081 cacctcccgc acccagttca acagcctgca gcagctcgtg gcttactact ccaaacatgc
     1141 tgatggcctg tgtcaccgcc tcactaccgt atgtcatcca ggctgaggag tggtactttg
     1201 gcaagatcac tagacgggaa tcagagcggc tgctgctcaa cgccgagaac ccgagaggga
     1261 ccttcctcgt gagggagagt gagaccacaa aaggtgccta ctgcctctct gtatccgact
     1321 tcgacaatgc caagggccta aatgtgaaac actacaagat ccgcaagctg gacagcggcg
     1381 gtttctacat cacctcccgc acccagttca acagcctgca gcagctcgtg gcttactact
     1441 ccaaacatgc tgatggcctg tgtcaccgcc tcactaccgt atgtctctag aactatagtg
     1501 agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     1561 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1621 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     1681 aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg
     1741 cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     1801 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1861 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1921 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1981 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2041 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2101 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2161 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2221 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2281 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2341 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2401 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2461 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2521 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2581 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2641 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2701 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2761 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2821 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2881 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2941 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3001 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3061 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3121 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3181 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3241 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3301 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3361 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3421 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3481 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3541 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3601 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3661 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3721 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3781 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3841 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3901 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     3961 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4021 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4081 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4141 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4201 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4261 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4321 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4381 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     4441 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccatagc caattcaata
     4501 tggcgtatat ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata
     4561 tggcgtatat ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata
     4621 tggcggactt ggcaccatgc caattcaata tggcggactt ggcactgtgc caactgggga
     4681 ggggtctact tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc
     4741 catattgaat tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat
     4801 atataggcaa tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta
     4861 tgtattggcc ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc
     4921 ccctcccaat gggcggtccc atataccata tatggggctt cctaataccg cccatagcca
     4981 ctcccccatt gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg
     5041 gtcctattga cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct
     5101 ggctcaatgc ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca
     5161 ttgcccattc atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac
     5221 ttggcagtac atcaatatct attaatagta acttggcaag tacattacta ttggaaggac
     5281 gccagggtac attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc
     5341 tgccaagtac atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca
     5401 ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta
     5461 gggaac
//
