LOCUS       sequence-9417-ex        4457 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 4457)
  AUTHORS   Jin M, Smith C, Hsieh HY, Gibson DF, Tait JF
  TITLE     Essential role of B-helix calcium binding sites in annexin 
            V-membrane binding
  JOURNAL   J Biol Chem 2004; 279:40351-57.
  PUBMED    15280367
REFERENCE   2  (bases 1 to 4457)
  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"; journalName: "J Biol 
            Chem"; date: "2004"; volume: " 279"; pages: "40351-57"
COMMENT     Sequence Label: pJ128
FEATURES             Location/Qualifiers
     source          1..4457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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     188..210
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     RBS             249..271
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(join(1345..1350,1351..1367,1368..1373))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      1543..1629
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1721..1748
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1802..1820)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1816..1863
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1900..2355
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1987..2006)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2197..2218
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2470..3261,3262..3330))
                     /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     3093..3112
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3423..4011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3912..3931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4165..4182
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4410..4429
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gacttacaat
       61 ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtacg ggcctcttcg
      121 ctattacgcc agcttgcgaa cggtgggtgc gctgcaaggc gattaagttg ggtaacgcca
      181 ggattctccc agtcacgacg ttgtaaaacg acggccagcg agagatcttg attggctagc
      241 agaataattt tgtttaactt taagaaggag atatagatat ggcgggtggt tgtggtcacg
      301 cacaggttct cagaggcact gtgactgact tccctggatt tgatgagcgg gctgatgcag
      361 aaactcttcg gaaggctatg aaaggcttgg gcacagatga ggagagcatc ctgactctgt
      421 tgacatcccg aagtaatgct cagcgccagg aaatctctgc agcttttaag actctgtttg
      481 gcagggatct tctggatgac ctgaaatcag aactaactgg aaaatttgaa aaattaattg
      541 tggctctgat gaaaccctct cggctttatg atgcttatga actgaaacat gccttgaagg
      601 gagctggaac aaatgaaaaa gtactgacag aaattattgc ttcaaggaca cctgaagaac
      661 tgagagccat caaacaagtt tatgaagaag aatatggctc aagcctggaa gatgacgtgg
      721 tgggggacac ttcagggtac taccagcgga tgttggtggt tctccttcag gctaacagag
      781 accctgatgc tggaattgat gaagctcaag ttgaacaaga tgctcaggct ttatttcagg
      841 ctggagaact taaatggggg acagatgaag aaaagtttat caccatcttt ggaacacgaa
      901 gtgtgtctca tttgagaaag gtgtttgaca agtacatgac tatatcagga tttcaaattg
      961 aggaaaccat tgaccgcgag acttctggca atttagagca actactcctt gctgttgtga
     1021 aatctattcg aagtatacct gcctaccttg cagagaccct ctattatgct atgaagggag
     1081 ctgggacaga tgatcatacc ctcatcagag tcatggtttc caggagtgag attgatctgt
     1141 ttaacatcag gaaggagttt aggaagaatt ttgccacctc tctttattcc atgattaagg
     1201 gagatacatc tggggactat aagaaagctc ttctgctgct ctctggagaa gatgactaaa
     1261 tcgaattcct gcccgggcgt tgtaatcata gtcataatca atactcctga ctgcgttagc
     1321 aatttaactg tgataaacta ccgcattaaa gctattcgat gataagctgt caaacatgat
     1381 aattcttgaa gacgaaaggg cctaggctga taaaacagaa tttgcctggc ggcagtagcg
     1441 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     1501 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     1561 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     1621 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     1681 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat
     1741 ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta
     1801 tccgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1861 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     1921 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1981 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2041 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2101 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2161 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2221 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2281 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2341 taacgtttac aatttctggc ggcacgatgg catgagatta tcaaaaagga tcttcaccta
     2401 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2461 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2521 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2581 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2641 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2701 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2761 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2821 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2881 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2941 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3001 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3061 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3121 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3181 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3241 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3301 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatcat gaccaaaatc
     3361 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3421 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3481 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3541 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3601 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3661 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3721 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3781 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3841 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3901 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3961 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4021 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4081 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4141 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagccgg cgataatggc
     4201 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     4261 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     4321 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     4381 gacagtcata agtgcggcga cgaccggtga attgtgagcg ctcacaattc tcgtgacatc
     4441 ataacgtccc gcgaaat
//
