LOCUS       Exported                5621 bp ds-DNA     circular SYN 06-MAR-2023
DEFINITION  pUC57.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5621)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 6, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     B-intron_kozak_R gtggcaagctttacgctgagaccagctcaccgagaaatga
            use with 2362 mTag-sGFP-F 
            ctcagcgtaaagcttgccaccatggTGTCCGAACTTATCAAAG
            Also use pUCR and pUCR-F for Nebuilder reaction
FEATURES             Location/Qualifiers
     source          1..5621
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(239..307)
                     /locus_tag="LacZ alpha"
                     /label=LacZ alpha
     primer_bind     364..386
                     /locus_tag="m13/pUC Forward"
                     /label=m13/pUC Forward
     primer_bind     378..395
                     /locus_tag="M13-fwd"
                     /label=M13-fwd
     misc_feature    432..479
                     /locus_tag="48bp_FRT3"
                     /label=48bp_FRT3
     misc_feature    480..499
                     /locus_tag="sgH (human ampka1 guide)"
                     /label=sgH (human ampka1 guide)
     misc_feature    510..529
                     /locus_tag="sgM site"
                     /label=sgM site
     misc_feature    533..580
                     /locus_tag="48bp_FRT"
                     /label=48bp_FRT
     misc_feature    727..732
                     /locus_tag="Kozak"
                     /label=Kozak
     misc_feature    730
                     /locus_tag="Vertebrate Kozak"
                     /label=Vertebrate Kozak
     misc_feature    733..1434
                     /locus_tag="zf_mTagBFP2 (+2ndVal)"
                     /label=zf_mTagBFP2 (+2ndVal)
     misc_feature    733..1434
                     /locus_tag="zf_mTagBFP2 (+2ndVal) noTAA"
                     /label=zf_mTagBFP2 (+2ndVal) noTAA
     misc_feature    733..736
                     /locus_tag="Vertebrate Kozak(1)"
                     /label=Vertebrate Kozak
                     /label=Vertebrate Kozak(1)
     misc_feature    733..735
                     /locus_tag="Kozak(1)"
                     /label=Kozak
                     /label=Kozak(1)
     variation       1435..1449
                     /locus_tag="(GGGGS)linker"
                     /label=(GGGGS)linker
     misc_feature    1450..1491
                     /locus_tag="V5-tag zf-optim (GKPIPNPLLGLDST)"
                     /label=V5-tag zf-optim (GKPIPNPLLGLDST)
     misc_feature    1501..3375
                     /locus_tag="synCoTC"
                     /label=synCoTC
     misc_feature    1501..1585
                     /locus_tag="afp 3'UTR (truncated at polyA)"
                     /label=afp 3'UTR (truncated at polyA)
     terminator      2066..3375
                     /locus_tag="CoTC terminator element"
                     /label=CoTC terminator element
     primer_bind     complement(3396..3416)
                     /locus_tag="M13-rev"
                     /label=M13-rev
     primer_bind     complement(3413..3435)
                     /locus_tag="m13/pUC Reverse"
                     /label=m13/pUC Reverse
     misc_binding    complement(3422..3444)
                     /locus_tag="LacO"
                     /label=LacO
     rep_origin      complement(3784..4466)
                     /direction=LEFT
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     misc_feature    complement(4561..5421)
                     /locus_tag="AmpR gene"
                     /label=AmpR gene
     CDS             complement(4564..5223)
                     /locus_tag="AmpR"
                     /label=AmpR
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tgaagttcct attccgaagt tcctattctt caaatagtat aggaacttcg
      481 aagatcggcc actacattct ggcctaggag gctgctgcgg ttccagaggt gggaagttcc
      541 tattccgaag ttcctattct ctagaaagta taggaacttc aataaaagtt aatttaagcg
      601 ttcattacaa taaattcaag tgaatctgtt ttaagctagc ctttaacgtt tttaaattcc
      661 ttcaaacaaa ttggatcctc tttgcagttc ctcatttctc ggtgagctgg tctcagcgta
      721 aagcttgcca ccatggtgtc cgaacttatc aaagaaaata tgcatatgaa gctgtacatg
      781 gagggaactg tggacaacca ccacttcaag tgcacctctg agggagaggg aaagccttat
      841 gagggaaccc agaccatgcg catcaaggtg gtggagggag gacctctgcc ttttgctttt
      901 gacatcctgg ccacctcttt cctgtatgga agcaagacct tcatcaacca cacccaggga
      961 atccctgact tcttcaagca gtctttccct gagggattca cctgggaaag agtgaccacc
     1021 tacgaggatg gaggagtgct gaccgccacc caggacacct ctctccagga tggatgcctg
     1081 atctacaacg tgaagatcag aggagtgaac tttactagca acggacctgt gatgcaaaag
     1141 aagaccctgg gatgggaggc tttcactgag accctgtacc cagctgatgg aggactggag
     1201 ggaagaaatg acatggctct gaagctggtg ggaggatctc acctgatcgc caatgccaag
     1261 accacctaca gaagcaagaa gcctgccaag aacctgaaga tgcctggagt gtactatgtg
     1321 gactacagac tggagcgcat caaggaggcc aacaatgaga cctatgtgga gcagcatgag
     1381 gtggctgtgg ccagatattg tgacctccct agcaagctgg gacacaagct gaatggagga
     1441 ggaggatctg gaaagccaat cccaaaccct ctgctgggac tggacagcac ctaaactagt
     1501 attaagtctc agccaccgtt aactgaacat gtcaaaacct gtggagactg ttgagatttg
     1561 atgttctgaa aagataaagc ctataaataa aacaatactc aaatacaaaa ggtatctttt
     1621 ttaaaaagtt agtttactgc ttccttaggc tctttctaaa tcccacccac ctgaaggaca
     1681 tcaattatgg atttatcctt catgccttgt ataaatacag atgtgcatat gaatttttat
     1741 gggtttttta aaaatgtggt tcattctata tacataacaa ttactttcaa catacataag
     1801 cctacctgta tttgtgcatt atattacatg gtacagataa atagtcatgt gccccatgaa
     1861 cattttggtc aaagatagat tacagataca atggtggtcc cataagatta tgatgccata
     1921 tttttactgt acctcttctt gtgcctagat acacaaatac tgacttgtgt tataattgcc
     1981 tgcagtattc aataatatgg tgtatagatt tgtagcctac gagctataac ctatactata
     2041 tagccaagga gtgcaggagg gtatgccatc tagatttgtg tgagtacact gatgtttgaa
     2101 caataaaatc acctaatgat aggtttctca aaccatatcc cagtccttaa gcaacacatg
     2161 actaatggat tcaaccattt cccctgttcg ttcagattgc tttttctttt tgcccctaca
     2221 attttgttga taatactgtt ttttggggct tttgtttttg tttttttttt tttttttgag
     2281 acaagagtct cactctgtcg tcaggctgga gtgcaatggc gctatctcag ctcactgcaa
     2341 cctctgcctc ccaggttcaa gcgattctcc tgcctcagcc tcctgagtag ctgagattac
     2401 aggcatgcgc caccgcgccc ggctaatttt gtatttttag tagagacgca gtttcaccat
     2461 gttggtcagg ctggtcttga acacccgacc tcaggtgatc cacccgcctc agcttcccaa
     2521 agtgctggga ttaaaggcgt gagccaccac acctggccaa tttactgttt tataccatgc
     2581 ttttttgtgt gtattctagg ttccctaaag caaacatgca ttttaaattt tgatagaacc
     2641 agatgacata caaaagtgcc tgttttctct gcattattaa aactgaaggt aatcagtctt
     2701 ttatttctat gactccaact ggcaaaaaat atttgccatt ttaacttttt ttttttgaga
     2761 cggagtcaaa aggtgccagt tttctctgca ttattaaaac tgaaggtaat cagtctttta
     2821 tttctatgac tccaactggc aaaagatatt tgccacgtta actttttttt tttttttttt
     2881 gagacggagg ctcacactgt cacccaggct gaagtgcagt ggcatgatct cagctcactt
     2941 gcaacctccg cctcccaggt tcaagtgatt ctcctgcctc agcctcccaa ggagttggga
     3001 ttacaggcgc atacccggct tattttttgt atttttagta gaaacgggat ttcaccgtgt
     3061 tagccaggat ggtctcaatc tccagacttc gtgatcgccc acctcggcct cccaaagtgc
     3121 tgggattaca ggcgtgagcc accgcgcccg gccctcagta ggcgattttt atatgtggta
     3181 taaagtagaa ttcttacaga tggatacccg tttatgccag tgccatgact gaaaccctgc
     3241 ttgtccaact tgaaatatct tcaatgccca tctattcttc cttgatcttg ttttttcctg
     3301 tgataataca atagttttta ccagttaatc agttaagatg aggcttacca aatatatggg
     3361 agtactactg aacacgatat caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3421 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3481 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3541 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4201 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5521 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5581 attaacctat aaaaataggc gtatcacgag gccctttcgt c
//
