LOCUS       sequence-7337-ex        3228 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3228)
  AUTHORS   Feng T, Li Z, Jiang W, Breyer B, Zhou L, Cheng H, Haydon RC, 
            Ishikawa A, Joudeh MA, He TC
  TITLE     Increased efficiency of cloning large DNA fragments using a lower 
            copy number plasmid.
  JOURNAL   Biotechniques. 2002 May . 32(5):992, 994, 996 passim.
  PUBMED    12019795
REFERENCE   2  (bases 1 to 3228)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Biotechniques. 2002 May . 32(5):992, 994, 996 passim."
COMMENT     Sequence Label: pMOLUC
FEATURES             Location/Qualifiers
     source          1..3228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..54
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(487..506)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(488..506)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(513..529)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             780..971
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(965..987)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1073..1213
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1128..1147
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1228..1245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1399..1987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1479..1498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2158..2949,2950..3018))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2781..2800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3019..3123)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3191..3209
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggaaacagct atgaccatga ttacgccaag ctcgaaatta accctcacta aagggaacaa
       61 aagctggtac gaggacaggc tggagccatg gctggtgacc acgtcgtgga atgccttcga
      121 attcagcacc tgcacatggg acgtcgacct gaggtaatta taacccgggc cctatatatg
      181 gatccaattg caatgatcat catgacagat ctgcgcgcga tcgatatcag cgctttaaat
      241 ttgcgcatgc tagctatagt tctagaggta ccggttgtta acgttagccg gctacgtata
      301 ctccggaata ttaataggcc taggatgcat atggcggccg cctgcagctg gcgccatcga
      361 tacgcgtacg tcgcgaccgc ggacatgtac agagctcgag aagtactagt ggccacgtgg
      421 gccgtgcacc ttaagcttgc ctaatcggac gaaaaaatga ccatgattac gccaagctcc
      481 aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttacc cggatctgca
      541 tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc
      601 attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct
      661 cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct
      721 ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag
      781 tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa
      841 cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc
      901 ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg
      961 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     1021 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     1081 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     1141 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     1201 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1261 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1321 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1381 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1441 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1501 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1561 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1621 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1681 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1741 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1801 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     1861 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1921 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1981 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2041 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2101 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2161 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2221 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2281 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2341 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2401 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2461 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2521 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2581 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2641 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2701 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2761 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2821 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2881 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2941 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3001 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3061 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3121 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3181 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaatt
//
