LOCUS       sequence-456469-        7033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fluorescent hybrid of tPA for PALM imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7033)
  AUTHORS   Scalettar BA, Shaver D, Kaech S, Lochner JE
  TITLE     Super-resolution imaging of neuronal dense-core vesicles.
  JOURNAL   J Vis Exp. 2014 Jul 2;(89). doi: 10.3791/51394.
  PUBMED    25046659
REFERENCE   2  (bases 1 to 7033)
  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 Vis 
            Exp."; date: "2014-07-2"; volume: "(89). doi"; pages: " 
            10.3791/51394"
COMMENT     Sequence Label: tPA-Dendra2
FEATURES             Location/Qualifiers
     source          1..7033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      96..231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(265..282)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(265..281)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(274..296)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      494..949
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(581..600)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     791..812
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(966..985)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1085..1107
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1145..1163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1231..1335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1336..1404,1405..2196)
                     /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(1554..1573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2367..2955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2856..2875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3109..3126
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3243..3264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3279..3284,3285..3302,3303..3309)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3317..3333
                     /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)."
     primer_bind     3322..3344
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3341..3357
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3341..3357
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        3386..3765
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3766..3969
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3920..3940
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3966..3990
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4200..4218
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4200..4217
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4231..4250
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      6705..6714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    6718..6852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6755..6774)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6809..6828
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        join(6921..7033,1..245)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(6921..7033,1..79)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6944..6964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      121 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      181 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      241 gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
      301 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      361 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      421 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
      481 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
      541 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      601 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      661 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
      721 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      781 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
      841 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      901 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
      961 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     1021 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     1081 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     1141 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1201 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1261 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1321 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1381 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1441 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1501 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1561 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1621 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1681 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1741 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1801 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1861 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1921 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1981 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2041 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2101 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2161 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2221 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2281 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2341 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2401 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2461 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2521 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2581 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2641 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2701 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2761 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2821 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2881 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2941 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3001 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3061 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3121 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3181 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3241 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     3301 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg
     3361 attacgaatt aattcgagct cgcccgacat tgattattga ctagttatta atagtaatca
     3421 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     3481 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     3541 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     3601 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     3661 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     3721 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     3781 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     3841 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     3901 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     3961 agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac
     4021 ctccatagaa gacaccggga ccgatccagc ctccgcggcc gggaacggtg cattggaacg
     4081 cggattcccc gtgccaagag tgacgtaagt accgcctata gagtctatag gcccaccccc
     4141 ttggcttcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga
     4201 tttaggtgac actatagaat aacatccact ttgcctttct ctccacaggt gtccactccc
     4261 aggtccaact gcacctcggt tctatcgatt gccaccatga agggagagct gttgtgcgtc
     4321 ctgctgcttt gtggagtggc gttcacgttg cctgaccagg gaatacacag gaggttcaga
     4381 agaggagctc ggtcctacag agcgacctgc agagatgaac agactcagac aacttaccaa
     4441 cagcatcagt catggctacg tcccatgctc agaggcaatc gggtggaata ctgccggtgc
     4501 aacagcggcc tggcacagtg ccactcggtg cccgtccgaa gttgcagtga accgagatgc
     4561 ttcaatgggg gtacgtgtca gcaggccctg tatttctctg acttcgtctg ccagtgccct
     4621 gacggatttg ttgggaaacg ctgtgatata gataccagag caacctgctt cgagggccag
     4681 ggcatcacct acagaggcac atggagcaca gcagaaaatg gggctgaatg catcaactgg
     4741 aatagcagtg ccctgtccca gaagccctac agtgcaagga ggccaaatgc catcaagctg
     4801 ggccttggga accacaatta ctgcagaaac ccagaccgag acgtgaagcc ctggtgctat
     4861 gtctttaagg cagggaagta taccacggag ttctgcagca cgccggcttg ccctaaggga
     4921 ccaactgagg actgctatgt tggaaaaggt gtgacttacc gtggcaccca cagctttacc
     4981 acatccaagg cctcctgcct cccatggaat tccatgatcc tgataggcaa gacttacaca
     5041 gcgtggaggg ccaactccca ggcacttggc ctgggcagac acaattattg ccggaaccca
     5101 gatggggatg ccaaaccttg gtgccacgtg atgaaggacc gaaagctgac atgggaatat
     5161 tgcgacatgt ccccatgctc cacctgcggc ctgaggcaat acaaacagcc tcagtttcga
     5221 attaaaggag gactcttcac agacatcacc tcacaccctt ggcaggccgc catctttgtc
     5281 aagaacaaga ggtctccagg agagagattc ctgtgtggag gggtgctgat cagttcctgc
     5341 tgggtgctat ctgccgccca ctgctttgta gagaggtttc caccccatca tcttaaagtg
     5401 gtcttgggca gaacatacag agtggtccct ggaaaggagg agcagacatt cgagatcgaa
     5461 aagtacatag tccataagga atttgatgac gacacttatg acaatgacat cgcattactg
     5521 cagctgaggt cagattccag tcagtgtgcc caggagagca gttctgtcgg cactgcctgc
     5581 ctccctgacc ccgacgtaca gcttcctgac tggacagagt gtgagctttc tggctacggc
     5641 aagcatgagg catcctctcc tttcttctct gaccggctga aggaggctca cgtcagactg
     5701 tatccgtcca gccgctgtac ctcacagcat ctgtttaaca aaaccatcac gagcaacatg
     5761 ctgtgtgcag gagacacccg aactgggggc aaccaagacg tccatgacgc gtgccagggt
     5821 gactcaggag gccctctggt gtgcatgatc gataagcgga tgactttact gggcatcatc
     5881 agctggggcc tcggctgtgg gcagaaggac gtgccaggga tatacacaaa ggtcactaat
     5941 tacctgaact ggatccaaga caacatgaag caaggaggag gaagtggagg aggatcccgg
     6001 ggtaacctga tcaaggagga catgcgcgtg aaggtgcaca tggagggcaa cgtgaacggc
     6061 cacgccttcg tgatcgaggg cgagggcaag ggcaagccct acgagggcac ccagaccgcc
     6121 aacctgaccg tgaaggaggg cgcccccctg cccttcagct acgacatcct gaccaccgcc
     6181 gtgcactacg gcaaccgggt gttcaccaag taccccgagg acatccccga ctacttcaag
     6241 cagagcttcc ccgagggcta cagctgggag cgcaccatga ccttcgagga caagggcatc
     6301 tgcaccatcc gcagcgacat cagcctggag ggcgactgct tcttccagaa cgtgcgcttc
     6361 aagggcacca acttcccccc caacggcccc gtgatgcaga agaagaccct gaagtgggag
     6421 cccagcaccg agaagctgca cgtgcgcgac ggcctgctgg tgggcaacat caacatggcc
     6481 ctgctgctgg agggcggcgg ccactacctg tgcgacttca agaccaccta caaggccaag
     6541 aaggtggtgc agctgcccga cgcccacttc gtggaccacc gcatcgagat cctgggcaac
     6601 gacagcgact acaacaaggt gaagctgtac gagcacgccg tggcccgcta cagccccctg
     6661 cccagccagg tgtggttgat ctagagtcga cctgcagaag cttggccgcc atggcccaac
     6721 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6781 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6841 catgtctgga tcgggaatta attcggcgca gcaccatggc ctgaaataac ctctgaaaga
     6901 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     6961 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     7021 tagtcagcaa cca
//
