LOCUS       sequence-148-exp        4091 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 4091)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 4091)
  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: L3502
FEATURES             Location/Qualifiers
     source          1..4091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..41
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(111..127)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(225..253)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1001..1021
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1119..1135
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1205..1221)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1246..1265)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1247..1265)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1275..1292)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1275..1291)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1284..1306)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1433..1888
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1520..1539)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1730..1751
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1914..2018
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2019..2087,2088..2879)
                     /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(2237..2256)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3050..3638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3539..3558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3792..3809
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3926..3947
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3962..3967,3968..3985,3986..3992)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4000..4016
                     /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     4005..4027
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4024..4040
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4024..4040
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4059..4079
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4061..4079
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctccacc gcggtggcgg ccgctctaga actagtggat ccaccggttc catggctagc
       61 cacgtgacgc gtggatcccc cgggctgcag gaattcgata tcaagcttat cgataccgtc
      121 gacctcgagg gggggcccgg tacgatatac taactaacaa gaattatcta atttttcagg
      181 taccggtaga aaaaatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg
      241 ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg
      301 atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc
      361 catgggtaag tttaaacata tatatactaa ctaaccctga ttatttaaat tttcagccaa
      421 cacttgtcac tactctcact tatggtgttc aatgcttctc gagataccca gatcatatga
      481 aacagcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat
      541 ttttcaaaga tgacgggaac tacaagacac gtaagtttaa acagttcggt actaactaac
      601 catacatatt taaattttca ggtgctgaag tcaagtttga aggtgatacc cttgttaata
      661 gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg
      721 aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca
      781 aagttgtaag tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagaact
      841 tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa
      901 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat
      961 ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa
     1021 cagctgctgg gattacacat ggcatggatg aactatacaa agtaagtttt aacatttaat
     1081 catacgaaat gaaagagctc caccgcggtg gcggccgctc tagaactagt ggatccaccg
     1141 gttccatggc tagccacgtg acgcgtggat cccccgggct gcaggaattc gatatcaagc
     1201 ttatcgatac cgtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg
     1261 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     1321 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     1381 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     1441 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1501 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1561 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1621 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     1681 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1741 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     1801 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1861 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     1921 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1981 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2041 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2101 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2161 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2221 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2281 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2341 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2401 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2461 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2521 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     2581 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2641 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2701 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2761 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     2821 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2881 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2941 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3001 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3061 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3121 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3181 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3241 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3301 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3361 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3421 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3481 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3541 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3601 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3661 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3721 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3781 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     3841 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     3901 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     3961 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     4021 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
     4081 aacaaaagct g
//
