LOCUS       sequence-384093-        4387 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 4387)
  TITLE     Integrated Genomics
REFERENCE   2  (bases 1 to 4387)
  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: L4440gtwy
FEATURES             Location/Qualifiers
     source          1..4387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(368..1159,1160..1228))
                     /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     991..1010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1229..1333)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1359..1814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1496..1517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1708..1727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1941..1963
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1955..1972
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1956..1972
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1982..2000
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2025..2041
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2026..2042
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2057..2181
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2222..2527)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(2324..2343)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2869..3528)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3443..3462
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(3582..3588,3589..3606,3607..3612))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(3637..3761)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(3783..3801)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3825..3842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3996..4387,1..197))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4076..4095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
       61 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      121 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
      181 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
      241 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
      301 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
      361 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
      421 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
      481 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
      541 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
      601 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
      661 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
      721 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
      781 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
      841 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
      901 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
      961 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     1021 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     1081 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     1141 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     1201 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     1261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     1321 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     1381 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     1441 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     1501 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1561 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     1621 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1681 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     1741 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1801 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     1861 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     1921 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc
     1981 gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg tcgacggtat
     2041 cgataagctt gatatcacca ctttgtacaa gaaagctgaa cgagaaacgt aaaatgatat
     2101 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
     2161 acaacatatc cagtcactat ggtcgacctg cagactggct gtgtataagg gagcctgaca
     2221 tttatattcc ccagaacatc aggttaatgg cgtttttgat gtcattttcg cggtggctga
     2281 gatcagccac ttcttccccg ataacggaga ccggcacact ggccatatcg gtggtcatca
     2341 tgcgccagct ttcatccccg atatgcacca ccgggtaaag ttcacgggag actttatctg
     2401 acagcagacg tgcactggcc agggggatca ccatccgtcg cccgggcgtg tcaataatat
     2461 cactctgtac atccacaaac agacgataac ggctctctct tttataggtg taaaccttaa
     2521 actgcatttc accagcccct gttctcgtca gcaaaagagc cgttcatttc aataaaccgg
     2581 gcgacctcag ccatcccttc ctgattttcc gctttccagc gttcggcacg cagacgacgg
     2641 gcttcattct gcatggttgt gcttaccaga ccggagatat tgacatcata tatgccttga
     2701 gcaactgata gctgtcgctg tcaactgtca ctgtaatacg ctgcttcata gcatacctct
     2761 ttttgacata cttcgggtat acatatcagt atatattctt ataccgcaaa aatcagcgcg
     2821 caaatacgca tactgttatc tggcttttag taagccggat ccacgcgttt acgccccgcc
     2881 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     2941 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     3001 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     3061 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     3121 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     3181 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     3241 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     3301 tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     3361 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     3421 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     3481 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     3541 agctcctgaa aatctcgacg gatcctaact caaaatccac acattatacg agccggaagc
     3601 ataaagtgta aagcctgggg tgcctaatgc ggccgccata gtgactggat atgttgtgtt
     3661 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
     3721 tcattttacg tttctcgttc agcttttttg tacaaacttg tgatatcaga tctgccggtc
     3781 tccctatagt gagtcgtatt aatttcgata agccaggttg cttcctcgct cactgactcg
     3841 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3901 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3961 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4021 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4081 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4141 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4201 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4261 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4321 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4381 gtaggcg
//
