LOCUS       sequence-60631-e        8042 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 8042)
  AUTHORS   Martell JD, Deerinck TJ, Sancak Y, Poulos TL, Mootha VK, Sosinsky 
            GE, Ellisman MH, Ting AY
  TITLE     Engineered ascorbate peroxidase as a genetically encoded reporter 
            for electron microscopy.
  JOURNAL   Nat Biotechnol. 2012 Oct 21. doi: 10.1038/nbt.2375.
  PUBMED    23086203
REFERENCE   2  (bases 1 to 8042)
  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: "Nat 
            Biotechnol. 2012 Oct 21. doi: 10.1038/nbt.2375."
COMMENT     Sequence Label: pcDNA3 Connexin43-GFP-APEX
FEATURES             Location/Qualifiers
     source          1..8042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2090..2092,2093..2095,2096..2806)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2135..2156)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2396..2415)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2743..2764
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3636..3653)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3642..3866
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3912..4340
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3999..4018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4209..4230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4349..4369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4354..4683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4534..4669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4596..4615
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4750..5544
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4804..4823)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5414..5433
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5718..5839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5755..5774)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5809..5828
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5888..5904)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5888..5904)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5901..5923)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5912..5928
                     /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(5936..5942,5943..5960,5961..5966))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5981..6002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6119..6136)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6290..6875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6370..6389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7046..7837,7838..7906))
                     /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     7669..7688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7907..8011)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgggtga ctggagtgcc ttggggaaat tactggacaa
      961 ggtccaagcc tactccaccg ctggagggaa ggtgtggctg tcagtgctct tcatattcag
     1021 aatcctgctc ctggggacag ctgttgagtc agcttggggt gatgaacagt ctgcctttcg
     1081 ctgtaacact caacaacctg gctgcgaaaa cgtctgctat gacaagtcct tccccatctc
     1141 tcacgtgcgc ttctgggtcc ttcagatcat attcgtgtct gtgcccacac tcctgtactt
     1201 ggcccatgtg ttctatgtga tgaggaagga agagaagcta aacaagaaag aagaggagct
     1261 caaagtggcc cagactgacg gggtcaacgt ggagatgcac ctgaagcaga ttgaaatcaa
     1321 gaagttcaag tacgggattg aagagcacgg caaggtgaaa atgaggggcg gcttgctgag
     1381 aacctacatc atcagcatcc tcttcaagtc tgtcttcgag gtggccttcc tgctcatcca
     1441 gtggtacatc tatgggttca gcttgagcgc ggtctacacc tgcaagagag atccctgccc
     1501 ccaccaggta gactgcttcc tctcacgtcc cacggagaaa accatcttca tcatcttcat
     1561 gctggtggtg tccttggtgt ctctcgcttt gaacatcatt gagctcttct acgtcttctt
     1621 caagggcgtt aaggatcgcg tgaagggaag aagcgatcct taccacgcca ccactggccc
     1681 actgagccca tcaaaagact gcggatctcc aaaatacgcc tacttcaatg gctgctcctc
     1741 accaacggct ccactctcgc ctatgtctcc tcctgggtac aagctggtta ctggtgacag
     1801 aaacaattcc tcgtgccgca attacaacaa gcaagctagc gagcaaaact gggcgaacta
     1861 cagcgcagag caaaatcgca tggggcaggc cggaagcacc atctccaact cgcacgccca
     1921 gccgttcgat ttccccgacg acaaccagaa tgccaaaaaa gttgctgctg gacatgaact
     1981 ccagccatta gccatcgtgg accaacgacc ttccagcaga gccagcagcc gcgccagcag
     2041 caggcctcgg cctgatgacc tggagattgg atccgggggt ggtgggtcta tggtgagcaa
     2101 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2161 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2221 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2281 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2341 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2401 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2461 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2521 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggc
     2581 caacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2641 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2701 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2761 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggggc gcgccaaggg
     2821 ctcgggctcg acctcgggct cgggcagggg aaaatcttac ccaactgtta gtcccgatta
     2881 ccaggacgcc attgaaaagg ctaagaggaa gctcagaggt tttatcgctg agaagaaatg
     2941 cgctcctcta attctccgtt tggcattcca ctctgctggt acttttgatt ccaagacaaa
     3001 gactggtggt cctttcggaa caattaagca ccaagctgag cttgctcatg gtgctaacaa
     3061 cggtcttgat atcgcggtta ggctgttgga gcctattaag gagcaattcc ctattgtgag
     3121 ctatgctgat ttctaccagt tggctggtgt tgttgctgtt gagattaccg gtggacctaa
     3181 ggttcctttc caccctggta gggaggacaa gcctgagcca ccacctgagg gtcgcttgcc
     3241 tgatgccact aagggttctg accatttgag ggatgtgttt ggaaaggcta tggggcttag
     3301 tgatcaggac attgttgctc tatctggtgg tcacaccatt ggagctgcac acaaggagcg
     3361 ttctggattt gagggaccgt ggacttctaa tcctctcatt tttgacaact catatttcac
     3421 tgagttgttg actggtgaga aggatggcct tcttcagttg ccaagtgata aggcactttt
     3481 gactgactct gtattccgcc ctcttgttga gaaatatgct gcggatgaag atgttttctt
     3541 tgctgattat gctgaagcac atcttaagct ctctgagctt ggatttgctg aagcctaact
     3601 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3661 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3721 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3781 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3841 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3901 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4321 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4501 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4621 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4681 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4741 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4801 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4861 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4921 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4981 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5041 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5101 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5161 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5221 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5281 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5341 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5401 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5461 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5521 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5581 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5641 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5701 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5761 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5821 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5881 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5941 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6001 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6061 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6121 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6181 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6241 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6301 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6361 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6421 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6481 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6541 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6601 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6661 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6721 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6781 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8041 tc
//
