LOCUS       Exported                7360 bp ds-DNA     circular SYN 09-AUG-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7360)
  AUTHORS   zhigang he lab
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 24, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7360
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     3412..3429
                     /label=Primer 3
                     /note="color: black; sequence: GTGTCTCAACCACTGTGA; added: 
                     2022-08-09"
     primer_bind     complement(3517..3535)
                     /label=Primer 2
                     /note="color: black; sequence: TCTTTGTAGCTGGCCATAG; added: 
                     2022-08-09"
     primer_bind     3536..3560
                     /label=Primer 1
                     /note="color: black; sequence: AGGAGAGGACAGTTTCTTTCCTTTG; 
                     added: 2022-08-09"
     protein_bind    3714..3743
                     /label=grade insertion
                     /bound_moiety="retro"
                     /note="color: #993366; direction: RIGHT"
     promoter        complement(4495..4513)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(4534..4550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    4558..4574
                     /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)."
                     /note="color: #31849b"
     promoter        complement(4582..4612)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 4582 .. 4588 / #ffffff / -10
                        2: 4589 .. 4606 / #ffffff
                        3: 4607 .. 4612 / #ffffff / -35"
     protein_bind    4627..4648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(4936..5524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5695..6555)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5695 .. 6486 / #ccffcc
                        2: 6487 .. 6555 / #ccffcc / signal sequence
                     Cleavage site after base 6486"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6556..6660)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(6686..7141)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     7283..7299
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        7309..7327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 gtcgacggta tcgggggagc tcgcagggtc tccattttga agcgggaggt ttgaacgcgc
       61 agccgccatg ccggggtttt acgagattgt gattaaggtc cccagcgacc ttgacgagca
      121 tctgcccggc atttctgaca gctttgtgaa ctgggtggcc gagaaggaat gggagttgcc
      181 gccagattct gacatggatc tgaatctgat tgagcaggca cccctgaccg tggccgagaa
      241 gctgcagcgc gactttctga cggaatggcg ccgtgtgagt aaggccccgg aggctctttt
      301 ctttgtgcaa tttgagaagg gagagagcta cttccacatg cacgtgctcg tggaaaccac
      361 cggggtgaaa tccatggttt tgggacgttt cctgagtcag attcgcgaaa aactgattca
      421 gagaatttac cgcgggatcg agccgacttt gccaaactgg ttcgcggtca caaagaccag
      481 aaatggcgcc ggaggcggga acaaggtggt ggatgagtgc tacatcccca attacttgct
      541 ccccaaaacc cagcctgagc tccagtgggc gtggactaat atggaacagt atttaagcgc
      601 ctgtttgaat ctcacggagc gtaaacggtt ggtggcgcag catctgacgc acgtgtcgca
      661 gacgcaggag cagaacaaag agaatcagaa tcccaattct gatgcgccgg tgatcagatc
      721 aaaaacttca gccaggtaca tggagctggt cgggtggctc gtggacaagg ggattacctc
      781 ggagaagcag tggatccagg aggaccaggc ctcatacatc tccttcaatg cggcctccaa
      841 ctcgcggtcc caaatcaagg ctgccttgga caatgcggga aagattatga gcctgactaa
      901 aaccgccccc gactacctgg tgggccagca gcccgtggag gacatttcca gcaatcggat
      961 ttataaaatt ttggaactaa acgggtacga tccccaatat gcggcttccg tctttctggg
     1021 atgggccacg aaaaagttcg gcaagaggaa caccatctgg ctgtttgggc ctgcaactac
     1081 cgggaagacc aacatcgcgg aggccatagc ccacactgtg cccttctacg ggtgcgtaaa
     1141 ctggaccaat gagaactttc ccttcaacga ctgtgtcgac aagatggtga tctggtggga
     1201 ggaggggaag atgaccgcca aggtcgtgga gtcggccaaa gccattctcg gaggaagcaa
     1261 ggtgcgcgtg gaccagaaat gcaagtcctc ggcccagata gacccgactc ccgtgatcgt
     1321 cacctccaac accaacatgt gcgccgtgat tgacgggaac tcaacgacct tcgaacacca
     1381 gcagccgttg caagaccgga tgttcaaatt tgaactcacc cgccgtctgg atcatgactt
     1441 tgggaaggtc accaagcagg aagtcaaaga ctttttccgg tgggcaaagg atcacgtggt
     1501 tgaggtggag catgaattct acgtcaaaaa gggtggagcc aagaaaagac ccgcccccag
     1561 tgacgcagat ataagtgagc ccaaacgggt gcgcgagtca gttgcgcagc catcgacgtc
     1621 agacgcggaa gcttcgatca actacgcgga caggtaccaa aacaaatgtt ctcgtcacgt
     1681 gggcatgaat ctgatgctgt ttccctgcag acaatgcgag agactgaatc agaattcaaa
     1741 tatctgcttc actcacggtg tcaaagactg tttagagtgc tttcccgtgt cagaatctca
     1801 acccgtttct gtcgtcaaaa aggcgtatca gaaactgtgc tacattcatc acatcatggg
     1861 aaaggtgcca gacgcttgca ctgcttgcga cctggtcaat gtggacttgg atgactgtgt
     1921 ttctgaacaa taaatgactt aaaccaggta tggctgccga tggttatctt ccagattggc
     1981 tcgaggacaa ccttagtgaa ggaattcgcg agtggtgggc tttgaaacct ggagcccctc
     2041 aacccaaggc aaatcaacaa catcaagaca acgctcgagg tcttgtgctt ccgggttaca
     2101 aataccttgg acccggcaac ggactcgaca agggggagcc ggtcaacgca gcagacgcgg
     2161 cggccctcga gcacgacaag gcctacgacc agcagctcaa ggccggagac aacccgtacc
     2221 tcaagtacaa ccacgccgac gccgagttcc aggagcggct caaagaagat acgtcttttg
     2281 ggggcaacct cgggcgagca gtcttccagg ccaaaaagag gcttcttgaa cctcttggtc
     2341 tggttgagga agcggctaag acggctcctg gaaagaagag gcctgtagag cagtctcctc
     2401 aggaaccgga ctcctccgcg ggtattggca aatcgggtgc acagcccgct aaaaagagac
     2461 tcaatttcgg tcagactggc gacacagagt cagtcccaga ccctcaacca atcggagaac
     2521 ctcccgcagc cccctcaggt gtgggatctc ttacaatggc ttcaggtggt ggcgcaccag
     2581 tggcagacaa taacgaaggt gccgatggag tgggtagttc ctcgggaaat tggcattgcg
     2641 attcccaatg gctgggggac agagtcatca ccaccagcac ccgaacctgg gccctgccca
     2701 cctacaacaa tcacctctac aagcaaatct ccaacagcac atctggagga tcttcaaatg
     2761 acaacgccta cttcggctac agcaccccct gggggtattt tgacttcaac agattccact
     2821 gccacttctc accacgtgac tggcagcgac tcatcaacaa caactgggga ttccggccta
     2881 agcgactcaa cttcaagctc ttcaacattc aggtcaaaga ggttacggac aacaatggag
     2941 tcaagaccat cgccaataac cttaccagca cggtccaggt cttcacggac tcagactatc
     3001 agctcccgta cgtgctcggg tcggctcacg agggctgcct cccgccgttc ccagcggacg
     3061 ttttcatgat tcctcagtac gggtatctga cgcttaatga tggaagccag gccgtgggtc
     3121 gttcgtcctt ttactgcctg gaatatttcc cgtcgcaaat gctaagaacg ggtaacaact
     3181 tccagttcag ctacgagttt gagaacgtac ctttccatag cagctacgct cacagccaaa
     3241 gcctggaccg actaatgaat ccactcatcg accaatactt gtactatctc tcaaagacta
     3301 ttaacggttc tggacagaat caacaaacgc taaaattcag tgtggccgga cccagcaaca
     3361 tggctgtcca gggaagaaac tacatacctg gacccagcta ccgacaacaa cgtgtctcaa
     3421 ccactgtgac tcaaaacaac aacagcgaat ttgcttggcc tggagcttct tcttgggctc
     3481 tcaatggacg taatagcttg atgaatcctg gacctgctat ggccagccac aaagaaggag
     3541 aggaccgttt ctttcctttg tctggatctt taatttttgg caaacaagga actggaagag
     3601 acaacgtgga tgcggacaaa gtcatgataa ccaacgaaga agaaattaaa actactaacc
     3661 cggtagcaac ggagtcctat ggacaagtgg ccacaaacca ccagagtgcc caactagcag
     3721 accaagacta cacaaaaact gctgcacagg cgcagaccgg ctgggttcaa aaccaaggaa
     3781 tacttccggg tatggtttgg caggacagag atgtgtacct gcaaggaccc atttgggcca
     3841 aaattcctca cacggacggc aactttcacc cttctccgct gatgggaggg tttggaatga
     3901 agcacccgcc tcctcagatc ctcatcaaaa acacacctgt acctgcggat cctccaacgg
     3961 ccttcaacaa ggacaagctg aactctttca tcacccagta ttctactggc caagtcagcg
     4021 tggagatcga gtgggagctg cagaaggaaa acagcaagcg ctggaacccg gagatccagt
     4081 acacttccaa ctattacaag tctaataatg ttgaatttgc tgttaatact gaaggtgtat
     4141 atagtgaacc ccgccccatt ggcaccagat acctgactcg taatctgtaa ttgcttgtta
     4201 atcaataaac cgtttaattc gtttcagttg aactttggtc tctgcgaagg gcgaattcgt
     4261 ttaaacctgc aggactagag gtcctgtatt agaggtcacg tgagtgtttt gcgacatttt
     4321 gcgacaccat gtggtcacgc tgggtattta agcccgagtg agcacgcagg gtctccattt
     4381 tgaagcggga ggtttgaacg cgcagccgcc aagccgaatt ctgcagatat ccatcacact
     4441 ggcggccgct cgactagagc ggccgccacc gcggtggagc tccagctttt gttcccttta
     4501 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4561 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4621 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4681 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4741 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4801 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4861 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4921 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4981 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5041 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5101 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5161 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5221 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5281 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5341 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5401 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5461 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5521 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5581 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5641 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5701 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5761 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5821 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5881 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5941 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6001 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6061 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6121 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6181 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6241 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6301 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6361 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6421 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6481 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6541 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6601 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6661 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     6721 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     6781 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     6841 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     6901 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     6961 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     7021 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     7081 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     7141 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7201 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     7261 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca
     7321 ctatagggcg aattgggtac cgggcccccc ctcgatcgag
//
