LOCUS       sequence-116004-        7940 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 7940)
  TITLE     pET23-CBD
REFERENCE   2  (bases 1 to 7940)
  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"
COMMENT     Sequence Label: pGFP(C3)-Vinculin
FEATURES             Location/Qualifiers
     source          1..7940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1333
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MGKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(650..678)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1273..1293
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    4735..4856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4772..4791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4826..4845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4863..5318)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5000..5021)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5212..5231
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5345..5449
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5451..5808
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5451..5642
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5474..5494)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5659..5794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5843..6637
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5843..6637
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5897..5916)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6507..6526
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6825..6844)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6869..6916
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7245..7833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7734..7753
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tgggtaaagg agaagaactt ttcactggag ttgtcccaat
      661 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
      721 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
      781 tgttccatgg ccaacacttg tcactacttt ctcttatggt gttcaatgct tttcaagata
      841 cccagatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca
      901 ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt
      961 tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg
     1021 aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc
     1081 agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg
     1141 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     1201 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     1261 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     1321 tgaactatac aagtactcag atctcgagct caagcttcga attcctgccg cgatgccagt
     1381 gtttcatacg cgtacgatcg agagcatcct ggagccggtg gcgcagcaga tctcgcacct
     1441 ggtgattatg cacgaggagg gcgaggtgga cggcaaagcc attcctgacc tcaccgcgcc
     1501 cgtagccgcc gtgcaggcgg ccgtcagcaa cctcgtccgg gttggaaaag agactgttca
     1561 gaccactgag gatcagattc tgaagagaga tatgccacca gcctttatta aggttgaaaa
     1621 tgcttgcacc aagcttgttc aggcagccca gatgcttcag tcagacccat actcggttcc
     1681 tgcgcgggat tacctcattg acggctctag gggaatcctt tctggcacat ctgacctact
     1741 gcttaccttc gatgaggctg aggttcgtaa aattattagg gtttgcaaag gaattttgga
     1801 atatcttaca gtggcagagg tagtggaaac tatggaagac ttggtcactt acacaaagaa
     1861 tcttgggcca ggaatgacta agatggccaa aatgattgat gagagacagc aggagttgac
     1921 tcaccaggaa caccgtgtga tgttggtgaa ctcaatgaac actgtcaaag agctgcttcc
     1981 tgttctcatt tcagctatga agatttttgt tacaaccaaa aactcaaaaa accaaggaat
     2041 agaagaagct ttgaaaaatc gaaattttac tgtagaaaag atgagtgctg aaattaacga
     2101 gatcattcgt gtgttacaac tcacttcctg ggatgaagat gcctgggcca gcaaggacac
     2161 tgaagccatg aagagagcgc tggcgtccat agactccaaa ttgaaccagg ccaaaggttg
     2221 gctccgtgac cccaatgcct ccccagggga tgctggagag caggccatca ggcagatctt
     2281 agatgaagct ggaaaagttg gtgaactttg tgcaggcaag gaacgcaggg agatcctagg
     2341 aacctgcaaa atgctagggc agatgactga ccaagtggct gacctccgag ccagaggaca
     2401 aggagcttcc ccagtggcca tgcagaaggc ccagcaagtg tctcaggggc tcgacgtgct
     2461 taccgccaaa gtggagaatg cagctcggaa gctggaagcc atgacgaact caaagcagag
     2521 cattgcaaag aagattgatg ctgcccagaa ttggctggcg gatccaaatg gtggacctga
     2581 gggagaagaa cagattcgag gggctttggc tgaagctcgg aagattgcag aattatgtga
     2641 tgatcctaag gagagagatg acatcctccg ctcccttgga gagatagctg ctctgacctc
     2701 taaactagga gacttgcgaa gacaggggaa aggagactcg ccagaggctc gagccttggc
     2761 taaacaagtg gcgacggcac tacagaacct gcagaccaaa accaacaggg ccgtggccaa
     2821 cagcagacct gccaaagcag ctgtccacct cgagggcaag attgaacagg cgcagcggtg
     2881 gattgataac cccacagtag atgaccgtgg agtcggtcag gctgccatcc gtggacttgt
     2941 ggctgagggg catcggctgg ccaatgtcat gatgggacct tatcgccaag atcttcttgc
     3001 caaatgtgac cgtgtagacc agctaacagc tcagctggct gacctggctg cccgagggga
     3061 gggggagagt cctcaggcga gagcacttgc atcccagctt caggactcct taaaggatct
     3121 taaagcccag atgcaggaag ctatgactca agaggtatcc gatgttttca gcgatactac
     3181 aactcctatc aagctgttgg cagtagccgc cactgctcct cctgatgcac ccaataggga
     3241 agaggtattt gatgaaaggg cagccaattc tgaaaaccat tcaggaaggc ttggagccac
     3301 agcagagaag gcggctgctg ttggtactgc caataaatcg acagtggaag gcattcaggc
     3361 atctgtgaag acagcccgag aactcactcc ccaggtcatc tccgctgctc ggatcttact
     3421 gaggaaccct ggtaaccagg ctgcttatga acattttgag accatgaaga accagtggat
     3481 tgataatgtt gaaaaaatga cagggctggt ggacgaggct attgatacca agtctctgtt
     3541 ggatgcttct gaagaagcaa ttaaaaaaga cctggacaag tgtaaggtag ccatggccaa
     3601 tattcagcct cagatgctgg tcgctggagc aaccagtatt gctcgtcggg ccaaccggat
     3661 tctgctggtt gctaagaggg aggtagagaa ctctgaggac ccgaagttcc gagaggctgt
     3721 gaaagctgcc tctgatgaac tgagcaaaac aatctccccc atggtgatgg atgccaaggc
     3781 tgtggctgga aacatctctg accctggcct gcaaaagagc ttcctggact caggatatcg
     3841 gatcctggga gctgaggcca aggtcagaga agccttccaa cctcaggagc ctgacttccc
     3901 gcctcctcca ccagaccttg aacagctacg actaactgat gagctggctc ctcctaagcc
     3961 acctctgcct gagggtgaag tccctccacc caggccccca ccaccagaag agaaggatga
     4021 agagttccct gagcagaaag ctggtgaggt gattaaccag ccaatgatga tggccgccag
     4081 gcagctccac gatgaagctc ggaaatggtc tagcaagggc aatgacatca ttgcagcagc
     4141 caagcgcatg gctctgctga tggcagagat gtctcggccg gtaagagggg gcagtggtac
     4201 caagcgggca cttattcagt gtgccaagga tatcgccaag gcctctgatg aggtgacgag
     4261 gttggccaag gaggttgcca agcagtgcac agataagcgg attagaacca atctcttaca
     4321 ggtatgcgag cgaatcccaa ctataagcac ccagctcaaa atcctatcca cagtgaaggc
     4381 cactatgctg ggccggacca acatcagtga tgaggagtct gagcaggcca cagagatgct
     4441 ggttcataat gcccagaacc tcatgcagtc tgtgaaggag actgtgcgag aggctgaagc
     4501 tgcttcaatc aaaatccgaa cagatgctgg ctttactctg cgctgggtca gaaagactcc
     4561 ctggtaccag tagggaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct
     4621 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     4681 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     4741 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4801 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     4861 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4921 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4981 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5041 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5101 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5161 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5221 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5281 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     5341 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5401 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     5461 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5521 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5581 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5641 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5701 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5761 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     5821 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5881 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5941 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     6001 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     6061 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     6121 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     6181 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     6241 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     6301 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6361 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6421 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6481 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6541 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6601 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     6661 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     6721 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     6781 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     6841 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     6901 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     6961 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     7021 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     7081 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7141 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7201 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7261 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7321 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7381 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7441 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7501 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7561 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7621 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7681 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7741 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7801 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7861 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7921 gtggataacc gtattaccgc
//
