LOCUS       157_pBlueScript         4957 bp    DNA     circular SYN 15-JAN-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4957)
  AUTHORS   NYU School of Medicine
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    695..1236
                     /label=Grb10 5' HA
     CDS             1252..1305
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(1306..1308,1309..2019)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     polyA_signal    2053..2174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2175..2714
                     /label=Grb10 3'HA
     promoter        complement(2769..2787)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2808..2824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2832..2848
                     /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(join(2856..2862,2863..2880,2881..2886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2901..2922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3210..3798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3969..4760,4761..4829))
                     /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"
     promoter        complement(4830..4934)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gctttagcag ctgcaatagc tgtattattc
      721 tgagggtgtc ttggactccc ctgatgaaca acccaaaaga tttttttcat gcttatttga
      781 taatatatag ctttttcagg aagcattgcc atcccaaatg gaataacttc attttgaact
      841 atacatgtag tatgtataca cacagactta acttgtattg tatgttacta atatgatttc
      901 atagtaatat taaaatacta tttggtttag gagcctgatc atccaaagat aaccattttg
      961 gtggaactct ataccctcac tgacattttt ttcttctctt gccatttttt atgtacagag
     1021 catggtcttg ggcttcaagt actaatgaac aacgatatta actcgtccgt ggaaagcctt
     1081 aactcagctt gcaacatgca gtctgatact gatactgcac cacttcttga ggatggccag
     1141 catgccagca accagggagc agcatctagc tcccggggac agccacaggc gtccccgagg
     1201 cagaaaatgc aacgctcgca gcctgtgcac attctcgaat tcggcagtgg agagggcaga
     1261 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag caagggcgag
     1321 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1381 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1441 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1501 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     1561 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1621 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1681 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1741 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1801 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1861 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1921 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1981 gccgccggga tcactctcgg catggacgag ctgtacaagg agttctaact agagctcgct
     2041 gatcagcctc gaaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2101 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2161 atcaatgtat cttaaggtgg gccagaacat gtgctgtctg cacagacaat gcgctctgct
     2221 ctccaaatgg tgatgggcag tgactgtaaa atcaagttag atgagtttgt tctcttaata
     2281 gcagctacag ttgttgccct ataatcttga ggaagccgac ccgcagtcct aaagccaaag
     2341 cacagtcctg ccccctaccc ttgtttactg tagttttccc tacacacaca gaacaatggt
     2401 gaattttggc ttattaatta gacactataa tatgtacaac aataattaat agtacaaaag
     2461 aacaattata atgctaatga agtcatatga gtgagttcta tccttcctgt tgtgatagtg
     2521 aagtgacaca aagtggtaag atggaaggaa gaaagtggga tagttgttgt gatacacaag
     2581 gctgctgttg accatctgtg gctatgtcat cagaattttc aagtcatgcc tctgccagct
     2641 gacctcaaag aactaaaagt gcagctctca atggctttgg gtatgctgtg taacatttca
     2701 gcgtttggtg tgaaggatcc actagttcta gagcggccgc caccgcggtg gagctccagc
     2761 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2821 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2881 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2941 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3001 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3061 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3121 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3181 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3241 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3301 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3361 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3421 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3481 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3541 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3601 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3661 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3721 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3781 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3841 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3901 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3961 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4021 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4081 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4141 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4201 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4261 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4321 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4381 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4441 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4501 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4561 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4621 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4681 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4741 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4801 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4861 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4921 ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
