LOCUS       sequence-123090-        5609 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 5609)
  AUTHORS   Burkel BM, von Dassow G, Bement WM
  TITLE     Versatile fluorescent probes for actin filaments based on the 
            actin-binding domain of utrophin.
  JOURNAL   Cell Motil Cytoskeleton. 2007 Nov . 64(11):822-32.
  PUBMED    17685442
REFERENCE   2  (bases 1 to 5609)
  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: "Cell Motil 
            Cytoskeleton. 2007 Nov . 64(11):822-32."
COMMENT     Sequence Label: GFP-UtrCH
FEATURES             Location/Qualifiers
     source          1..5609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(90..92,93..95,96..806)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(90..92,93..95,96..806)
                     /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(135..156)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(396..415)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     743..764
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1658..1792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1682..1701)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1736..1755
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1905..1925)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1905..1923)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1944..1960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1944..1960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1957..1979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1968..1984
                     /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(1992..1998,1999..2016,2017..2022))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2037..2058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2175..2192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2346..2934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2426..2445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3105..3896,3897..3965))
                     /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     3728..3747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3966..4070)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4096..4551)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4233..4254)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4445..4464
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4625..5609
                     /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 atccaaacca tggtgagcaa gggcgaggag ctgttcaccg
      121 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      181 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      241 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
      301 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      361 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      421 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      481 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
      541 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
      601 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      661 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
      721 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
      781 ctctcggcat ggacgagctg tacaagtccg gaaccatggc caagtatgga gaacatgaag
      841 ccagtcctga caatgggcag aacgaattca gtgatatcat taagtccaga tctgatgaac
      901 acaatgacgt acagaagaaa acctttacca aatggataaa tgctcgattt tcaaagagtg
      961 ggaaaccacc catcaatgat atgttcacag acctcaaaga tggaaggaag ctattggatc
     1021 ttctagaagg cctcacagga acatcactgc caaaggaacg tggttccaca agggtacatg
     1081 ccttaaataa cgtcaacaga gtgctgcagg ttttacatca gaacaatgtg gaattagtga
     1141 atataggggg aaccgacatt gtggatggaa atcacaaact gactttgggg ttactttgga
     1201 gcatcatttt gcactggcag gtgaaagatg tcatgaagga tgtcatgtcg gacctgcagc
     1261 agacgaacag tgagaagatc ctgctcagct gggtgcgtca gaccaccagg ccctacagcc
     1321 aagtcaacgt cctcaacttc accaccagct ggacagatgg actcgccttt aatgctgtcc
     1381 tccaccgaca taaacctgat ctcttcagct gggataaagt tgtcaaaatg tcaccaattg
     1441 agagacttga acatgccttc agcaaggctc aaacttattt gggaattgaa aagctgttag
     1501 atcctgaaga tgttgccgtt cggcttcctg acaagaaatc cataattatg tatttaacat
     1561 ctttgtttga ggtgctacct cagcaagtca ccatagacta actcgagctc aagcttcgaa
     1621 ttctgcagtc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
     1681 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1741 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1801 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     1861 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     1921 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1981 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2041 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2101 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2161 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2221 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2281 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2341 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2401 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2461 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2521 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2581 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2641 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2701 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2761 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2821 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2881 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2941 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3001 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3061 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3121 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3181 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3241 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3301 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3361 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3421 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3481 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3541 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3601 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3661 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3721 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3781 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3841 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3901 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3961 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4021 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4081 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4141 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4201 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4261 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4321 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4381 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4441 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4501 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     4561 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4621 agtcgaccat agccaattca atatggcgta tatggactca tgccaattca atatggtgga
     4681 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga
     4741 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga
     4801 cttggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg
     4861 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat
     4921 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg
     4981 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg
     5041 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg
     5101 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct
     5161 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt
     5221 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc
     5281 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac
     5341 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc
     5401 aagtacatta ctattggaag gacgccaggg tacattggca gtactcccat tgacgtcaat
     5461 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg
     5521 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg
     5581 tctatataag caatgctcgt ttagggaac
//
