LOCUS       sequence-131-exp        3963 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 3963)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 3963)
  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: L2822
FEATURES             Location/Qualifiers
     source          1..3963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..35
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     19..35
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     54..74
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        56..74
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    87..194
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     111..127
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(161..177)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(235..263)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1011..1031
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    1089..1196
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1113..1129
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1163..1179)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1204..1223)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1205..1223)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1233..1250)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1233..1249)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1242..1264)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1391..1846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1478..1497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1688..1709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1872..1976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1977..2045,2046..2837)
                     /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(2195..2214)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3008..3596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3497..3516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3750..3767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3884..3905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3920..3925,3926..3943,3944..3950)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(3958..3963,1..11)
                     /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(3963,1..22)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
       61 accctcacta aagggaacaa aagctggagc tccaccgcgg tggcggccgc tctagaacta
      121 gtggatcccc cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagg
      181 gggggcccgg taccggtaga aaaaatgagt aaaggagaag aacttttcac tggagttgtc
      241 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
      301 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
      361 ctacctgttc catgggtaag tttaaacata tatatactaa ctaaccctga ttatttaaat
      421 tttcagccaa cacttgtcac tactttctgt tatggtgttc aatgcttctc gagataccca
      481 gatcatatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
      541 agaactatat ttttcaaaga tgacgggaac tacaagacac gtaagtttaa acagttcggt
      601 actaactaac catacatatt taaattttca ggtgctgaag tcaagtttga aggtgatacc
      661 cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga
      721 cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag
      781 aatggaatca aagttgtaag tttaaacatg attttactaa ctaactaatc tgatttaaat
      841 tttcagaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat
      901 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
      961 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
     1021 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa atgcccgggg
     1081 gatcggtgga gctccaccgc ggtggcggcc gctctagaac tagtggatcc cccgggctgc
     1141 aggaattcga tatcaagctt atcgataccg tcgacctcga gggggggccc ggtacccaat
     1201 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     1261 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     1321 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1381 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1441 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1501 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1561 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1621 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1681 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1741 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     1801 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     1861 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1921 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1981 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2041 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2101 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2161 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2221 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2281 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2341 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2401 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2461 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2521 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2581 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2641 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2701 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2761 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2821 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2881 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2941 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3001 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3061 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3121 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3781 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3841 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     3901 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     3961 tga
//
