LOCUS       sequence-256-exp        5248 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 5248)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 5248)
  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: L4271
FEATURES             Location/Qualifiers
     source          1..5248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..27
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11..27
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        48..66
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             95..808
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(818..834)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     847..863
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1631..1647)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1660..1676
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1705..2421)
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2436..2452)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2484..2503)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2485..2503)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2510..2527)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2510..2526)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2519..2541)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2668..3123
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2755..2774)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2965..2986
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3149..3253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3254..3322,3323..4114)
                     /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(3472..3491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4285..4873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4774..4793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5027..5044
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5161..5182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5197..5202,5203..5220,5221..5227)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(5235..5248,1..3)
                     /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     join(5240..5248,1..14)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caatttcaca caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac
       61 taaagggaac aaaagctggg taccggtaga aaaaatgagt aaaggagaag aacttttcac
      121 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
      181 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
      241 tactggaaaa ctacctgttc catggccaac acttgtcact actttctctt atggtgttca
      301 atgcttttca agatacccag atcatatgaa acggcatgac tttttcaaga gtgccatgcc
      361 cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg
      421 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
      481 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
      541 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca
      601 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg
      661 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
      721 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
      781 tacacatggc atggatgaac tatacaaatg cccgggggat ccactagttc tagagcggcc
      841 ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcggaagat gggttccatg
      901 cgccaaagtt aaggatacga aggctcatat tgatggtctc aagaagggac aaacatatca
      961 gttccgtgtc aaggctgtca ataaggaagg agcttctgat gcattgtcta ctgataagga
     1021 caccaaagcc aagaatccat atgatgagcc aggaaaaacc ggaactccgg atgttgtcga
     1081 ctgggatgct gatcgtgtct cacttgaatg ggaaccacca aagtctgacg gaggagctcc
     1141 aatcactcaa tacgtcattg agaagaaggg caaacatgga agagactggc aagaatgcgg
     1201 aaaggtttct ggagatcaaa ccaatgctga gattcttgga ctcaaggagg gagaagagta
     1261 ccagttccgt gtgaaggctg ttaacaaggc cggaccggga gaggcttcag acccaagccg
     1321 aaaggttgtt gcaaagccaa gaaacttgaa gccatggatt gatcgtgaag caatgaagac
     1381 gatcactatc aaggtaggaa acgatgtgga attcgatgtt ccagtacgcg gagaaccacc
     1441 accgaagaag gaatggatct tcaatgagaa accagtcgat gatcaaaaga tcaggattga
     1501 aagcgaagac tacaagaccc gatttgtgct ccgtggagca actcgcaagc atgctggttt
     1561 gtacactctt actgctacca acgcttctgg aagcgacaaa cattccgttg aggtcattgt
     1621 gctcgggggg gatccactag ttctagagcg gccccccctc gaggtcgacg gtatcgataa
     1681 gcttgatatc gaattctacg aatgctattt gtatagttca tccatgccat gtgtaatccc
     1741 agcagctgtt acaaactcaa gaaggaccat gtggtctctc ttttcgttgg gatctttcga
     1801 aagggcagat tgtgtggaca ggtaatggtt gtctggtaaa aggacagggc catcgccaat
     1861 tggagtattt tgttgataat ggtctgctag ttgaacgctt ccatcttcaa tgttgtgtct
     1921 aattttgaag ttaactttga ttccattctt ttgtttgtct gccatgatgt atacattgtg
     1981 tgagttatag ttgtattcca atttgtgtcc aagaatgttt ccatcttctt taaaatcaat
     2041 accttttaac tcgattctat taacaagggt atcaccttca aacttgactt cagcacgtgt
     2101 cttgtagttc ccgtcatctt tgaaaaatat agttctttcc tgtacataac cttcgggcat
     2161 ggcactcttg aaaaagtcat gccgtttcat atgatctggg tatcttgaaa agcattgaac
     2221 accataagag aaagtagtga caagtgttgg ccatggaaca ggtagttttc cagtagtgca
     2281 aataaattta agggtaagtt ttccgtatgt tgcatcacct tcaccctctc cactgacaga
     2341 aaatttgtgc ccattaacat caccatctaa ttcaacaaga attgggacaa ctccagtgaa
     2401 aagttcttct cctttactca ttttttctac cgggggatcc actagttcta gagcggccgc
     2461 caccgcggtg gagctccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg
     2521 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     2581 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     2641 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     2701 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2761 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2821 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2881 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2941 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3001 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3061 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     3121 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3181 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3241 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3301 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3361 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3421 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3481 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3541 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3601 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3661 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3721 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3781 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3841 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3901 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3961 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4021 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4081 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4141 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4201 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4261 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4321 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4381 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     4441 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4501 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4561 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4621 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4681 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4741 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4801 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4861 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4921 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4981 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5041 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5101 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5161 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     5221 tatgttgtgt ggaattgtga gcggataa
//
