LOCUS       sequence-200377-        5354 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 5354)
  AUTHORS   Pantoja R, Rodriguez EA, Dibas MI, Dougherty DA, Lester HA
  TITLE     Single-molecule imaging of a fluorescent unnatural amino acid 
            incorporated into nicotinic receptors.
  JOURNAL   Biophys J. 2009 Jan . 96(1):226-37.
  PUBMED    19134478
REFERENCE   2  (bases 1 to 5354)
  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: "Biophys J. 
            2009 Jan . 96(1):226-37."
COMMENT     Sequence Label: pAMV:nAChRgamma-eGFP
FEATURES             Location/Qualifiers
     source          1..5354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    129..150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(165..170,171..188,189..195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    203..219
                     /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     208..230
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     227..243
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     227..243
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        264..282
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(join(900..1610,1611..1613))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(900..1610,1611..1613))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(942..963)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1291..1310
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1550..1571
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2806..2825)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2807..2825)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2832..2849)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2832..2848)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2841..2863)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2990..3445
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3077..3096)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3287..3308
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3471..3575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3576..3644,3645..4436)
                     /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(3794..3813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4607..5195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5096..5115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(5349..5354,1..12)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
       61 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
      121 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
      181 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
      241 gaccatgatt acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggtaccg
      301 cggccgcttt tttttttttt ttttttttcc cctctgtctc tttatttctc tatcttctcc
      361 caggaagcct tgggtatttc cccaagagca cactcagcat caacacggga tacagctcta
      421 caaacagcat ggccctcata gctgactcca gggcccagga gggattggtt ggctcagtct
      481 ggcaaaggca ggtaggggcg ggggtctccg gggaacggca ggtcaggcac ttggttgtag
      541 tgggccatga ggaagatgcc agcagtgcca cagatgaaga gggagagcat ggctaggaag
      601 cagactcggt ccagcactcg gcccaccagc aaccactcct cgttcccact gtcaaagtga
      661 ctctgctggc gtcgggcacg agccatgagg ttacaggcat ccacacaggc ctggatggct
      721 ggggaggctt gcttcaggct gccacagaac tcctggctct gcctcacttc tggaccattc
      781 tctagcttct ccaatactgc ctgcactaat ccattgcgct gcctttgcct aaagaggagt
      841 tcgcttcgag gcagacagag gtccccttcc tctcgagcca tgatgggcca ccctgaggac
      901 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
      961 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
     1021 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     1081 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
     1141 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     1201 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     1261 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     1321 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     1381 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
     1441 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
     1501 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     1561 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct catacctgag
     1621 gaagagccat tctggagtcg gaaccgggca tcctggacag cagctggagc tagtgggcgc
     1681 acatgcatcc gtaacagctg gggcaggagc ctcaagaaca acttgcggac cccacgggcc
     1741 atggagtgtg tgtgggggga ccgcaaggac acattgagca cgaccacaga gttcacgacg
     1801 atgaggatgg tcaccaccat gaggaaggtc aggtacttgc tgatgagtgg cactgcctgg
     1861 gaggtctcag gcaccttctt agccacaagg aaaaggaaga cagtctgggc caggagcacg
     1921 ttggtggcca ctgtgcattt ctggccgccc gccttagcag gaaggaagta gatgaggatg
     1981 gcgactgagg agatgagcac acagggggcg atgatgttga tgacgtagaa gaggggcttg
     2041 cgctggataa gcaggtagaa caccaccttc tggtggcccg cctcctctgc aggagccacg
     2101 gagtccagga gcattttagc cggtcggtgc cggatggccc actccccatt ctctgtgaaa
     2161 gcctccgggt caatgaagat ccactcaatg gcttgcccat cctcctggct cagctgcaag
     2221 ttgatctcac tggtgctgta agtctgggat tggaagatga gggaacagtt ctgccaatcg
     2281 aaggggaagt aggtgacaga gatggagcag gaggagcgga agatggcagg cggcagccag
     2341 tagatacaac cgtcggggga cacgaggaca ttgcagtaga gagccacctc gaagacaccg
     2401 tccacattgt tctccaggac gatatctggc cgccagacca tagtagatgg caccctcaat
     2461 atccacaggc cttcgtagtc ttttgggtcc cagcgcaggc gatagtcgca ccattgcatc
     2521 tctatccaga cgttagttgt gagggcctcc tctcgttcat tcagggagat gaggttggtc
     2581 aaggtaagct tcaggctgac attgaccaca tctgagtcgc gctcagccgg ccgcaggtgg
     2641 gggtcgtagt ttcgcatcag gtccgcaagc agacgctcct cttggttgcg gctctgggcc
     2701 cccaggcaga cagccaacag cagcaagagg agatgaggtc tctggccccc ttgcatggtg
     2761 gaagtatttg aaagaaaatt aaaaataaaa acgagctcca attcgcccta tagtgagtcg
     2821 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2881 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2941 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     3001 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3061 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3121 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3181 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3241 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3301 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3361 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3421 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3541 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3601 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3661 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3721 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3781 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3841 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3901 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3961 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4021 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4081 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4141 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4201 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4261 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4321 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4381 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4441 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4741 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5221 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5281 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5341 ccgagcgcag cgag
//
