LOCUS       sequence-450116-        5522 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 5522)
  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 5522)
  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 
            Neurosci."; date: "2005-08-17"; volume: "25(33)"; pages: "7669-81"
COMMENT     Sequence Label: pCS2-myc-gfp-dSH2
FEATURES             Location/Qualifiers
     source          1..5522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..1001
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1036..1054
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1036..1053
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1100..1129
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1130..1840
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1157..1185)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1780..1800
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    2575..2709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2599..2618)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2653..2672
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2822..2842)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2822..2840)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2861..2877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2861..2877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2874..2896)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2885..2901
                     /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(2909..2915,2916..2933,2934..2939))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2954..2975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3092..3109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3263..3851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3343..3362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4022..4813,4814..4882))
                     /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     4645..4664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4883..4987)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5013..5468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5150..5171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5362..5381
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact catgccaatt
       61 caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt
      121 caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt
      181 caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag
      241 tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat
      301 aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc
      361 tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta
      421 ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata
      481 ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc
      541 tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc
      601 ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata
      661 ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg
      721 ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa
      781 tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc
      841 attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc
      901 aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg
      961 cctaatggga ggtctatata agcaatgctc gtttagggaa ccgccattct gcctggggac
     1021 gtcggagcaa gcttgattta ggtgacacta tagaatacaa gctacttgtt ctttttgcag
     1081 gatcgatccc atcgatatgg agcaaaagct tattagcgag gaagatctga gtaaaggaga
     1141 agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
     1201 caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa
     1261 atttatttgc actactggaa aactacctgt tccatggccc acacttgtca ctactttcac
     1321 ttatggtgtt caatgctttt caagataccc agatcatatg aaacggcatg actttttcaa
     1381 gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa
     1441 ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
     1501 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
     1561 taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt
     1621 caaaattaga cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa
     1681 tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc
     1741 tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac
     1801 agctgctggg attacacatg gcatggatga actatacaag aatttcgagc tcaagcttaa
     1861 gagcaactat gtggcgccct ccgactccat ccaggctgaa gagtggtact ttgggaagat
     1921 cactcgtcgg gagtccgagc ggctgctgct caaccccgaa aacccccggg gaaccttctt
     1981 ggtccgggag agcgagacga caaaaggtgc ctattgcctc tccgtttctg actttgacaa
     2041 cgccaagggg ctcaatgtga agcactacaa gatccgcaag ctggacagcg gcggcttcta
     2101 catcacctca cgcacacagt tcagcagcct gcagcagctg gtggcctact actccaaaca
     2161 tgctgatggg ctgtgtcacc gcctcactac caagcttagc aactatgtgg cgccctccga
     2221 ctccatccag gctgaagagt ggtactttgg gaagatcact cgtcgggagt ccgagcggct
     2281 gctgctcaac cccgaaaacc cccggggaac cttcttggtc cgggagagcg agacgacaaa
     2341 aggtgcctat tgcctctccg tttctgactt tgacaacgcc aaggggctca atgtgaagca
     2401 ctacaagatc cgcaagctgg acagcggcgg cttctacatc acctcacgca cacagttcag
     2461 cagcctgcag cagctggtgg cctactactc caaacatgct gatggcctgt gtcaccgcct
     2521 cactacctaa tctagatgag gatccgctag aactatagtg agtcgtatta cgtagatcca
     2581 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2641 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2701 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2761 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
     2821 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2881 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
     2941 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3001 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3061 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3121 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3181 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3241 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3301 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3361 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3421 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3481 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3541 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3601 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3661 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3721 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3781 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3841 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3901 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3961 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4021 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4081 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4141 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4201 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4261 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4321 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4381 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4441 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4501 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4561 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4621 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4681 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4741 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4801 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4861 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4921 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4981 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     5041 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5101 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5161 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5221 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5281 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5341 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5401 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5461 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5521 ct
//
