LOCUS       sequence-295-exp        4676 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 4676)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4676)
  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: L4842
FEATURES             Location/Qualifiers
     source          1..4676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1202..1218)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1202..1218)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1255..1275)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1255..1273)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1294..1310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1294..1310)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1307..1329)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1318..1334
                     /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(1342..1348,1349..1366,1367..1372))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1387..1408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1525..1542)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1696..2284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1776..1795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2455..3246,3247..3315))
                     /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     3078..3097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3316..3420)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3446..3901)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3583..3604)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3795..3814
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4028..4050
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4042..4059
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4043..4059
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4069..4088
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4069..4087
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tcgaggatgc caagagacgc caagcccaaa agatcaacga gctccaagag gctcttgatg
       61 ccgccaactc taagaacgct tctcttgaga agaccaagtc cagacttgtc ggagacttgg
      121 acgacgctca agttgatgtc gagagagcca acggtgtcgc cagtgctctt gagaagaagc
      181 aaaagggctt cgacaagatc atcgacgaat ggagaaagaa gaccgacgac ttggccgctg
      241 agcttgatgg agctcaacgc gaccttcgca acacctccac tgatctcttc aaggccaaga
      301 acgcccaaga ggagctcgcc gaggttgttg agggactccg ccgtgagaac aagagcttga
      361 gccaagagat caaggatctt accgatcaac tcggagaggg aggacgctct gtccacgaaa
      421 tgcaaaagat catccgccgt cttgagattg agaaggaaga actccaacac gctttggacg
      481 aggctgaggc tgcccttgaa gctgaagaga gcaaggttct ccgcgcccag gttgaagttt
      541 cccagatccg ttccgaaatc gagaaacgca tccaggagaa ggaggaagag ttcgagaaca
      601 cgagaaagaa ccacgcccgc gctcttgaat caatgcaagc ttggcgagat tttcaggagc
      661 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
      721 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
      781 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
      841 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
      901 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
      961 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     1021 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     1081 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     1141 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tggaaccggt
     1201 ggatccacta gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta
     1261 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1321 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1381 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1441 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1501 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1561 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1621 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1681 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1741 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1801 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1861 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1921 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1981 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2041 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2101 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     2161 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2221 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2281 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2341 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2401 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2461 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2521 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2581 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2641 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2701 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2761 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2821 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2881 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2941 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3001 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3061 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3421 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     3481 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3541 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3601 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3661 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3721 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3781 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3841 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3901 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     3961 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     4021 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca
     4081 ctatagggcg aattgggtac ccggttccat ggtgaaaacg ggggcgaaga agttgtccat
     4141 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     4201 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     4261 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     4321 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     4381 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     4441 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     4501 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     4561 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     4621 ctccatttta gcttccttag ctcctgaaaa tctcgccaag cttgcatgcc tgcagg
//
