LOCUS       Exported                2963 bp ds-DNA     circular SYN 19-JUL-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Oct 28, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2963
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2653..2661
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /label=source:synthetic DNA construct(1)
                     /note="/ApEinfo_hidden="
                     /note="/locus_tag=source:synthetic DNA construct(1)"
     source          2662..2676
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /note="/ApEinfo_hidden="
                     /note="/locus_tag=source:synthetic DNA construct"
     source          2677..2684
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /label=source:synthetic DNA construct(2)
                     /note="/ApEinfo_hidden="
                     /note="/locus_tag=source:synthetic DNA construct(2)"
     primer_bind     complement(12..29)
                     /locus_tag="L4440"
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      183..771
                     /direction=RIGHT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /locus_tag="pBR322ori-F"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(942..1802)
                     /codon_start=1
                     /gene="bla"
                     /locus_tag="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1565..1584
                     /locus_tag="Amp-R"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
     primer_bind     1975..1993
                     /locus_tag="pBRforEco"
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     primer_bind     complement(2031..2053)
                     /locus_tag="pGEX 3'"
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2148..2403)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /locus_tag="lacZ-alpha"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MFTGRRFTTS*LGKPWRYPT*SPCSTSPFRQLA**RRGPHRSPFP
                     TVAQPEWRMAPDAVFSPYASVRYFTPHMVHSQYNLL*CRI"
     primer_bind     2153..2172
                     /locus_tag="pRS-marker"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     2366..2388
                     /locus_tag="M13/pUC Forward"
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2380..2397
                     /locus_tag="M13 Forward"
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
     primer_bind     2381..2397
                     /locus_tag="M13 fwd"
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(2398..2403)
                     /locus_tag="MCS"
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    2404..2652
                     /locus_tag="U6"
                     /label=U6
     misc_feature    2650..2652
                     /locus_tag="AgeI\(ACCGGT)"
                     /label=AgeI\(ACCGGT)
                     /note="/vntifkey=21"
     misc_feature    2653
                     /locus_tag="U6 TSS"
                     /label=U6 TSS
     misc_feature    2685..2720
                     /locus_tag="PspCas13b DR"
                     /label=PspCas13b DR
     misc_feature    2721..2725
                     /locus_tag="T5"
                     /label=T5
     CDS             complement(2726..2748)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /locus_tag="lacZ-alpha(1)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /label=lacZ-alpha(1)
                     /translation="MTMITPS"
     misc_feature    complement(2726..2731)
                     /locus_tag="MCS(1)"
                     /label=MCS
                     /label=MCS(1)
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2744..2760)
                     /locus_tag="M13 rev"
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2744..2760)
                     /locus_tag="M13 Reverse"
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2757..2779)
                     /locus_tag="M13/pUC Reverse"
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    complement(2768..2784)
                     /locus_tag="lac operator"
                     /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(2792..2822)
                     /locus_tag="lac promoter"
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2837..2858)
                     /locus_tag="CAP binding site"
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2041 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2101 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2161 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     2221 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2281 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2341 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     2401 ttcgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
     2461 gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
     2521 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
     2581 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
     2641 ggacgaaaca ccgatgtctt cgatattcaa gcgtcggaag acaagttgtg gaaggtccag
     2701 ttttgagggg ctattacaac tttttaagct tggcgtaatc atggtcatag ctgtttcctg
     2761 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2821 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2881 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2941 gaggcggttt gcgtattggg cgc
//
