LOCUS       060_PBS-N-mCTCFh        4904 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 4904)
  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..4904
                     /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"
     primer_bind     678..697
                     /label=223
     primer_bind     complement(678..697)
                     /label=192
     misc_feature    698..1197
                     /label=L hom
     CDS             1195..2661
                     /codon_start=1
                     /translation="MKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPE
                     AAAFVKVSMDGAPYLRKIDLRMYKASMVSKGEELFTGVVPILVELDGDVNGHKFSVSGE
                     GEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEG
                     YVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV
                     YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSK
                     DPNEKRDHMVLLEFVTAAGITLGMDELYKGAGAQASNSAVDEGEAVEAIVEESETFIKG
                     KERKTYQRRREGGQEEDACHLPQNQTDGGEVVQDVNSSVQMVMMEQLDPTLLQMKTEVM
                     EGTVAPEAEAAVDDTQIITLQVVNMEEQPINIGELQLVQVPVPVTVPVATTSVEELQGA
                     YENEVSKEGLAESEPMICHTLPLPEGFQVVK"
     misc_feature    1198..1401
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
     primer_bind     1361..1380
                     /label=p242
     misc_feature    join(1408..1410,1411..1413,1414..2124)
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
     primer_bind     complement(1882..1900)
                     /label=p243
     primer_bind     complement(2097..2113)
                     /label=EGFP-CR
     misc_feature    2161..2661
                     /label=R hom
     primer_bind     complement(2662..2681)
                     /label=224
     primer_bind     2662..2681
                     /label=191
     primer_bind     complement(2663..2679)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2716..2734)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2755..2771)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2779..2795
                     /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(2803..2809,2810..2827,2828..2833))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2848..2869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3157..3745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3916..4707,4708..4776))
                     /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(4777..4881)
                     /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 gcttgataac caaaattaac attgtagtgt
      721 ttctcttaga ctgaacttct gcagtatatt taacacctgc tgtcaccagt tttctgttaa
      781 ttttgagtat aatctagagt cttgatcagt ttcagatttt tttgttttga ttggcaagat
      841 ttataatata ggtggtgaat ttttttttct tccatcaaag aggaagatat ctattaaata
      901 accattgatg gatgcctaat tcattcacta aggtgggttt tttggggggt caggaggttg
      961 agttttaggg gtttggcttg gtttttggtt cttagagata gggtttctct gtgcaggcca
     1021 ggctggcctc aaactcagaa atctgcctgc ctgtgggttc taagtgctgg gattaaaggt
     1081 gtgtgcacca ctaccacctg ccactcaaat gattttagtt taaaatgtgt gctttgattt
     1141 gagcaaacca catcataaat aacatctgtg ttctccttaa taaaggaagg ggagatgaag
     1201 gagaagagtg cttgtcctaa agatccagcc aaacctccgg ccaaggcaca agttgtggga
     1261 tggccaccgg tgagatcata ccggaagaac gtgatggttt cctgccaaaa atcaagcggt
     1321 ggcccggagg cggcggcgtt cgtgaaggta tcaatggacg gagcaccgta cttgaggaaa
     1381 atcgatttga ggatgtataa agctagcatg gtgagcaagg gcgaggagct gttcaccggg
     1441 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1501 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1561 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     1621 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1681 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1741 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1801 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1861 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1921 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1981 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     2041 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2101 ctcggcatgg acgagctgta caagggtgca ggcgcccaag cttcgaattc tgcagtcgac
     2161 gaaggtgaag ccgtagaggc gatagtcgag gagtctgaaa ctttcattaa aggaaaagaa
     2221 agaaagactt accagagacg ccgggaaggg ggccaggaag aagatgcttg ccacctgccc
     2281 cagaaccaga cagatggggg tgaggtggtc caggatgtca acagcagtgt acagatggta
     2341 atgatggaac agctggatcc tacccttctc cagatgaaga ctgaagtcat ggagggtaca
     2401 gtggctccgg aagcagaggc tgcagtggac gatacccaga tcataacctt gcaggttgta
     2461 aatatggagg aacagcccat taacatagga gagcttcagc ttgtccaagt acctgttcct
     2521 gtgacggtac ctgttgctac tacttcagta gaagaacttc agggggctta tgagaatgaa
     2581 gtgtctaaag agggccttgc agaaagtgaa ccgatgatat gtcacacctt acctttgcct
     2641 gaaggatttc aggtggtgaa aggatccact agttctagag cggccgccac cgcggtggag
     2701 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2761 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2821 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2881 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2941 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3001 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3061 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3121 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3181 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3241 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3301 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3361 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3421 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3481 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3541 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3601 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3661 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3721 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3781 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3841 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3901 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3961 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4021 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4081 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4141 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4201 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4261 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4321 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4381 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4441 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4501 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4561 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4621 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4681 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4741 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4801 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4861 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccac
//
